PQDx 0450-027-00 WHO PQ Public Report November 2021, version 3.

WHO Prequalification of In Vitro Diagnostics

PUBLIC REPORT

Product: Abbott RealTime HCV

WHO reference number: PQDx 0450-027-00

Abbott RealTime HCV with product codes 4J86-90, 4J86-80 and 4J86-70, manufactured by
Abbott Molecular Inc, CE-Marked regulatory version, was accepted for the WHO list of
prequalified in vitro diagnostics and was listed on 10 December 2019.

Summary of WHO prequalification assessment for Abbott RealTime HCV

Date Outcome
Prequalification listing 10 December 2019 listed
Dossier review N/A N/A
Site inspection(s) of quality 21 November 2019 MR
management system
Product performance Quarter (Q) 3 and Q4 of 2019 MR
evaluation
MR: Meet Requirements
N/A: Not Applicable

Report amendments and/or product changes

This public report has since been amended. Amendments may have arisen because of
changes to the prequalified product for which WHO has been notified and has undertaken
a review. Amendments to the report are summarized in the following table, and details of
each amendment are provided below.

Version Summary of amendment Date of report

amendment
2.0 The manufacturer has submitted a change request to extend the claim 16 September
of the Abbott RealTime HCV assay (4J86) to include quantification of 2020
hepatitis C viral RNA (HCV RNA) in whole blood spotted on cards
as dried blood spots (DBS) (i.e. obtained via venipuncture or capillary
blood) from HCV-infected individuals.
3.0 Updated Abbott’s European Authorized Representative (EC Rep) legal 1 November
entity name from Abbott GmbH & Co. KG to Abbott GmbH. Labeling 2021
changes to comply with the labeling requirements for product
registered under IVDR.

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PQDx 0450-027-00 WHO PQ Public Report November 2021, version 3.0

Intended use:
According to the claim of intended use from Abbott Molecular Inc, “Abbott RealTime HCV
assay is an in vitro reverse transcription polymerase chain reaction (RT‑ PCR) assay for the
quantitation of hepatitis C viral ribonucleic acid (HCV RNA) in human serum, plasma and
whole blood spotted on cards as dried blood spots (DBS) (ie, obtained via venipuncture or
capillary blood) from HCV-infected individuals. The Abbott RealTime HCV assay is intended
for use as an aid in the management of HCV-infected patients undergoing antiviral therapy.
The Abbott RealTime HCV assay is not for screening blood, plasma, serum or tissue donors
for HCV, or to be used as a diagnostic test to confirm the presence of HCV infection.

The intended users for the Abbott RealTime HCV assay are laboratory and healthcare
professionals.”

Assay description:
According to the claim of assay description from Abbott Molecular Inc, “Abbott RealTime
HCV assay consists of 3 reagent kits:
• Abbott RealTime HCV Amplification Reagent Kit
• Abbott RealTime HCV Control Kit
• Abbott RealTime HCV Calibrator Kit
The Abbott RealTime HCV assay uses RT-PCR to generate amplified product from the RNA
genome of HCV in clinical specimens. An RNA sequence that is unrelated to the HCV target
sequence is introduced into each specimen at the beginning of sample preparation. This
unrelated RNA sequence is simultaneously amplified by RT-PCR and serves as an internal
control (IC) to demonstrate that the process has proceeded correctly for each sample. The
amount of HCV target sequence that is present at each amplification cycle is measured
through the use of fluorescent‑labeled oligonucleotide probes on the Abbott m2000rt
instrument. The probes do not generate signal unless they are specifically bound to the
amplified product. The amplification cycle at which fluorescent signal is detected by the
Abbott m2000rt is proportional to the log of the HCV RNA concentration present in the
original sample”.

Test kit contents:

Component Number of tests and product code

Abbott RealTime HCV Amplification Reagent Kit 96 tests

(product code 4J86-90)

Abbott RealTime HCV Internal Control 4 vials x 1.2mL (4J86Y)

Abbott RealTime HCV 4 packs X 24 tests/pack (4J86)
Amplification Reagent Pack

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PQDx 0450-027-00 WHO PQ Public Report November 2021, version 3.0

Abbott RealTime HCV Control Kit product code 4J86-80

Abbott RealTime HCV Negative Control 8 vials x 1.8mL/ vial (4J86Z)
Abbott RealTime HCV Low Positive Control 8 vials x 1.3ml/vial (4J86W)

Abbott RealTime HCV High Positive Control 8 vials x 1.3mL/vial (4J86X)

Abbott RealTime HCV Calibrator Kit Product code 4J86-70

Abbott RealTime HCV Calibrator A 12 vials x 1.3 mL/vial (4J86A)
Abbott RealTime HCV Calibrator B 12 vials x 1.3 mL/vial (4J86B)

Items required but not provided:

Component Product code and description

Sample preparation area
Abbott m2000sp instrument 9K14-02
Abbott m1000sp instrument 4J72-01
Abbott m24sp instrument 3N06-01
Abbott RealTime HCV m2000 ROW System Combined 1L69-07 or higher
Application CD-ROM
Abbott m2000sp software with version 6.0 or higher 50-148400 or higher
5 mL Reaction Vessels General Lab material
Abbott mSample Preparation System 04J70-24
Calibrated precision pipettes 20 µL to 1 000 µL
Aerosol barrier pipette tips 20 µL to 1 000µL
13 to 16 mm sample tubes 13 to 16 mm sample tubes
200 μL disposable tips 04J71-17
1000 μL disposable tips 04J71-17
Vortex mixer General lab equipment
Abbott Optical Adhesive Covers 04J71-75
Abbott Adhesive Cover Applicator 9K32-01
Abbott Splash-Free Support Base 09K31-01
200 mL Reagent Vessels 4J71-60
Master Mix Tube 04J71-80
Abbott 96-Deep-Well Plate 04J71-30
Abbott 96-Well Optical Reaction Plate 04J71-70
Centrifuge capable of 2000g General lab equipment
1.4 mL Micro Vial 15 mm Caps (optional) 3N20-01
Amplification area

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PQDx 0450-027-00 WHO PQ Public Report November 2021, version 3.0

Abbott m2000rt 9K15-01

Abbott RealTime HCV m2000 ROW System Combined 1L69-07 or higher
Application CD-ROM
Abbott m2000rt Optical Calibration Kit List No. 4J71-93
Other Materials

Biological safety cabinet approved for working with General lab equipment
infectious materials
Sealable plastic bags General lab material
RNase-free water (Eppendorf or equivalent) † General lab material
1.7 mL molecular biology grade microcentrifuge tubes
(Dot Scientific, Inc. or equivalent) †
Cotton Tip Applicators (Puritan or equivalent) † General lab material

†NOTE: These 3 items are used in the procedure for Monitoring the Laboratory for the
Presence of Contamination. Refer to the QUALITY CONTROL PROCEDURES section of this
package insert.

Storage:

• The Abbott RealTime HCV Amplification Reagent Pack and Internal Control
vials(4J86-90) must be stored at – 10°C or colder when not in use. Care must be taken
to separate the Abbott RealTime HCV Amplification Reagent Pack that is in use from
direct contact with samples, calibrators, and controls.
• The Abbott RealTime HCV Negative and Positive Controls (4J86-80) must be stored
at – 10°C or colder.
• The Abbott RealTime HCV Calibrator A and Calibrator B (4J86-70) must be stored at
– 10°C or colder.
Shipping:

• Abbott RealTime HCV Amplification Reagent Kit: Ship on dry ice.

• Abbott RealTime HCV Control Kit: Ship on dry ice.
• Abbott RealTime HCV Calibrator Kit: Ship on dry ice.

Shelf-life upon manufacture:

18 months.

Warnings/limitations:

Refer to the current instructions for use.

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PQDx 0450-027-00 WHO PQ Public Report November 2021, version 3.0

Prioritization for prequalification

Based on the established eligibility criteria, Abbott RealTime HCV was given priority for WHO
prequalification assessment.

Product dossier assessment

In accordance with the WHO procedure for abridged prequalification assessment, Abbott
Molecular Inc. was not required to submit a product dossier for Abbott RealTime HCV as per
the “Instructions for compilation of a product dossier” (PQDx_018 version 3).
Notwithstanding, certain aspects of the product dossier previously submitted for stringent
regulatory review were reviewed by an assessor during the site inspection.

Commitments for prequalification:

Abbott Molecular Inc to include a table of numbers of specimens per analyte tested for
potentially interfering substances in the IFU by May 31, 2024.

Manufacturing site inspection

In accordance with the WHO procedure for abridged prequalification assessment, a desk
assessment was performed in lieu of an onsite inspection. The desk assessment covered
the site(s) of manufacture of Abbott RealTime HCV, as per the “Information for
manufacturers on prequalification inspection procedures for the sites of manufacture of
diagnostics” (PQDx_014 version 4).

The desk assessment found that the manufacturer had an acceptable quality management
system and good manufacturing practices in place that ensured the consistent manufacture
of a product of good quality.

The desk assessment was found to be acceptable on 21 November 2019 and that the quality
management system for Abbott RealTime HCV, meets WHO prequalification requirements.

Product performance evaluation

The Abbott RealTime HCV assay is an in vitro reverse transcription polymerase chain
reaction (RT‑PCR) assay for the quantitation of hepatitis C viral ribonucleic acid (HCV RNA)
in human serum and EDTA/ACD plasma from HCV-infected individuals. The Abbott RealTime
HCV assay is intended for use as an aid in the management of HCV-infected patients
undergoing antiviral therapy. A volume of 0.2 mL or 0.5 mL of specimen is used to perform
the assay (however, at least 0.7 – 1.3 mL of specimen should be available for use with the
m1000 and m2000sp systems). This type of assay requires laboratory equipment and cannot
be performed in laboratories with limited facilities.

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The Abbott RealTime HCV assay was evaluated at the National Serology Reference
Laboratory (NRL), Melbourne, Australia, on behalf of WHO in Q3 and Q4 2019. From this
evaluation, we drew the following conclusions.
Analytical evaluation
The precision (intra- and inter-assay) of measurement was found to be acceptable. At 102
IU/mL, CV% were <8% and at 104 IU/mL, CV% were < 3%.
The limit of detection of the assay was estimated at 31 IU/mL (95% CI: 11-52 IU/mL), which
is consistent with the claim (30 IU/mL with the 0.2 mL sample preparation procedure).
There was no cross-contamination when high positive and negative specimens were ran
together.
All 6 HCV genotypes included in the 4th HCV RNA Genotype Panel for Nucleic Acid
Amplification (NIBSC reference: 14/209) and in the NRL HCV Mixed Genotype Panel were
detected by the assay.
Clinical evaluation
In this limited performance evaluation on a panel of 197 ACD plasma specimens, using a
sample volume of 0.2 mL, we found a bias of -0.05 log10 IU/ mL (limits of agreement: -0.42 to
0.32 log10 IU/mL) compared to the reference results using the cobas HCV quantitative nucleic
acid test with cobas 6800 System (Roche Diagnostics GmbH). The sensitivity (N=99) was 100%
(95% CI: 96.3% – 100%) and specificity (N=98) was 100% (95% CI: 96.2% - 100%) compared
to the reference results.
In this study, the invalid rate was 0%.

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Labelling

1. Labels
2. Instructions for use

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PQDx 0450-027-00 WHO PQ Public Report November 2021, version 3.0

1. Labels

1.1 Abbott RealTime HCV Amplification Reagent Kit box label (4J86-90)

Page 8 of 32
 Producto de EE.UU. / Prodotto negli USA / Fabricado nos EUA
Abbott RealTime é uma marca comercial de Abbott Laboratories.
Product of USA / Produkt aus USA / Produit des Etats-Unis / Armored RNA é uma marca comercial registada de Ambion.
ProClin é uma marca comercial registada de Rohm and Haas.
• 1 Frasco (0,40 ml) de reagente de activação. Solução de cloreto de manganésio 30 mM. Conservantes: 0,1% de ProClin 300 e 0,15% de ProClin 950.
ProClin 300 e 0,15% de ProClin 950.
tamponada com um corante de referência. Conservantes: 0,1% de
• 1 Frasco (1,10 ml) de reagente de oligonucleótidos do HCV. <0,1% de oligonucleótidos sintéticos (4 primers e 2 sondas), e <0,3% de dNTPs numa solução
• 1 Frasco (0,141 ml) de enzima rTth da polimerase termoestável (2,9 a 3,5 Unidades/µl) numa solução tamponada.
contém:
2. Abbott RealTime HCV Amplification Reagent Pack (4 embalagens, 24 testes/embalagem). Cada embalagem de reagentes
do HIV, anticorpos anti‑HIV‑1/HIV-2 e anti-HCV. Conservantes: 0,1% de ProClin® 300 e 0,15% de ProClin 950.
sequências de controlo interno em plasma humano negativo. Plasma humano negativo testado e considerado não-reactivo para HBsAg, ADN do HBV, ARN do HCV, ARN
1. Abbott RealTime HCV Internal Control (4 frascos, 1,2 ml por frasco). <0,01% de Armored RNA® (ARN protegido) não-infeccioso com
Conteúdo:
rastreio do HCV em amostras de sangue, plasma, soro ou tecido de dadores, nem como teste de diagnóstico confirmatório da presença de infecção pelo HCV.
a ser utilizado como meio auxiliar no tratamento de doentes infectados pelo HCV a receber terapêutica anti-viral. O ensaio Abbott RealTime HCV não se destina a ser utilizado para o
para a quantificação do ácido ribonucleico viral da hepatite C (ARN do HCV) em soro e plasma humanos de indivíduos infectados pelo HCV. O ensaio Abbott RealTime HCV destina-se
(pt) Para utilização in vitro. O ensaio Abbott RealTime HCV é um ensaio, para utilização in vitro, de transcriptase reversa associada à reacção em cadeia da polimerase (RT-PCR)
(4x24 Tests)
HCV
Amplification Reagent Kit Abbott RealTime
de utilização.
materiais de origem humana como potencialmente infecciosos. Consultar as instruções
potenzialmente infettivi. Consultare le istruzioni per l’uso. / ATENÇÃO: manusear os
las instrucciones de uso. / ATTENZIONE: Trattare i materiali di origine umana come
maneje los productos de origen humano como potencialmente infecciosos. Consulte
étaient potentiellement infectieux. Consulter les instructions d’utilisation. / ATENCIÓN:
beachten. / ATTENTION : Manipuler les produits d’origine humaine comme s’ils
muss mit der entsprechenden Vorsicht gehandhabt werden. Gebrauchsanweisung
instructions for use. / ACHTUNG: Humanmaterial gilt als potentiell infektiös und
CAUTION: Handle human sourced materials as potentially infectious. Consult
Abbott RealTime is a trademark of Abbott Laboratories.
Armored RNA is a registered trademark of Ambion.
51-602125/R5 ProClin is a registered trademark of Rohm and Haas.
0.1% ProClin 300 and 0.15% ProClin 950.
• 1 Bottle (0.40 mL) Activation Reagent. 30 mM manganese chloride solution. Preservatives:
300 and 0.15% ProClin 950.
2 probes), and <0.3% dNTPs in a buffered solution with a reference dye. Preservatives: 0.1% ProClin
• 1 Bottle (1.10 mL) HCV Oligonucleotide Reagent. <0.1% synthetic oligonucleotides (4 primers and
1434 • 1 Bottle (0.141 mL) Thermostable rTth Polymerase Enzyme (2.9 to 3.5 Units/µL) in buffered solution.
pack). Each Reagent Pack contains:
2. Abbott RealTime HCV Amplification Reagent Pack (4 packs, 24 tests/
anti‑HIV‑1/HIV-2, and anti-HCV. Preservatives: 0.1% ProClin® 300 and 0.15% ProClin 950.
human plasma tested and found to be nonreactive for HBsAg, HBV DNA, HCV RNA, HIV RNA,
<0.01% noninfectious Armored RNA® with internal control sequences in negative human plasma. Negative
1. Abbott RealTime HCV Internal Control (4 vials, 1.2 mL per vial).
Contents:
of HCV infection.
screening blood, plasma, serum or tissue donors for HCV, or to be used as a diagnostic test to confirm the presence
P333+P313, P362+P364, P501
management of HCV-infected patients undergoing antiviral therapy. The Abbott RealTime HCV assay is not for
P261, P280, P272, P302+P352,
H317 and plasma from HCV-infected individuals. The Abbott RealTime HCV assay is intended for use as an aid in the
4J86-90 chain reaction (RT-PCR) assay for the quantitation of hepatitis C viral ribonucleic acid (HCV RNA) in human serum
(en) For In Vitro Diagnostic Use. The Abbott RealTime HCV assay is an in vitro reverse transcription-polymerase
(4x24 Tests) HCV
Amplification Reagent Kit Abbott RealTime
(de) In-vitro-Diagnostikum. Der Abbott RealTime HCV Assay ist ein In-vitro-Reverse-Transkription-Polymerasekettenreaktions-Assay (RT-PCR) zur (es) Para uso en diagnóstico in vitro. Abbott RealTime HCV es un ensayo in vitro de reacción en cadena de la polimerasa previa transcripción inversa
quantitativen Bestimmung von Hepatitis-C-viraler Ribonukleinsäure (HCV RNA) in Humanserum und -plasma von HCV-infizierten Personen. Der Abbott (RT-PCR) para la cuantificación del ácido ribonucleico del virus de la hepatitis C (RNA del VHC) en suero y plasma humanos de individuos infectados
RealTime HCV Assay dient als Hilfsmittel bei der Behandlung von HCV-infizierten Patienten unter antiviraler Therapie. Der Abbott RealTime HCV Assay con el VHC. El ensayo Abbott RealTime HCV se usa como ayuda en la gestión de la terapia antirretroviral en pacientes infectados con el VHC. El ensayo
ist nicht zum Screening von Blut-, Plasma-, Serum- oder Gewebespendern auf HCV oder zur Verwendung als diagnostischer Test zur Bestätigung einer Abbott RealTime HCV no debe utilizarse como análisis de cribado de VHC en donaciones de sangre, plasma, suero o de tejidos ni como análisis de
HCV-Infektion vorgesehen. diagnóstico para la confirmación de la infección por VHC.
Inhalt: Contenido:
1. Abbott RealTime HCV Interne Kontrolle (4 Fläschchen, 1,2 ml pro Fläschchen). <0,01 % nicht infektiöse 1. Control interno Abbott RealTime HCV (4 tubos de 1,2 ml cada uno). <0,01% Armored RNA® no infeccioso
Armored RNA® mit internen Kontrollsequenzen in negativem Humanplasma. Negatives Humanplasma wurde getestet und war nicht reaktiv con secuencias de control interno en plasma humano negativo. El plasma humano negativo se ha analizado y no se ha encontrado
für HBsAg, HBV DNA, HCV RNA, HIV RNA, anti‑HIV‑1/HIV-2 und anti-HCV. Konservierungsmittel: 0,1 % ProClin® 300 und 0,15 % ProClin reactividad para el HBsAg, ni para el DNA del VHB, ni el RNA del VHC, ni el RNA del VIH, ni reactividad de anticuerpos anti‑VIH‑1/VIH-2 ni
950. anti-VHC. Conservantes: ProClin® 300 al 0,1% y ProClin 950 al 0,15%.
2. Abbott RealTime HCV Amplifikations-Reagenzpackung (4 Packungen, 24 Tests/Packung). Jede 2. Envase de reactivo de amplificación Abbott RealTime HCV (4 envases de 24 análisis cada uno).
Reagenzpackung enthält: Cada envase de reactivo contiene:
• 1 Fläschchen (0,141 ml) Thermostabiles rTth-Polymerase-Enzym (2,9 bis 3,5 Einheiten/µl) in gepufferter Lösung. • 1 Frasco (0,141 ml) de Thermostable rTth Polymerase Enzyme (enzima polimerasa termoestable rTth) (2,9 a 3,5 unidades/µl) en
• 1 Fläschchen (1,10 ml) HCV Oligonukleotidreagenz. <0,1 % synthetische Oligonukleotide (4 Primer und 2 Sonden) und <0,3 % solución tamponada.
dNTPs in einer gepufferten Lösung mit einem Referenzfarbstoff. Konservierungsmittel: 0,1 % ProClin 300 und 0,15 % ProClin 950. • 1 Frasco (1,10 ml) de HCV Oligonucleotide Reagent (reactivo de oligonucleótidos del VHC). <0,1% synthetic oligonucleotides
• 1 Fläschchen (0,40 ml) Aktivierungsreagenz. 30 mmol/l Manganchloridlösung. Konservierungsmittel: 0,1 % ProClin 300 und 0,15 % (oligonucleótidos sintéticos) (4 cebadores y 2 sondas) y <0,3% dNTPs en una solución tamponada con un colorante de referencia.
ProClin 950. Conservantes: ProClin 300 al 0,1% y ProClin 950 al 0,15%.
ProClin ist ein eingetragenes Warenzeichen von Rohm and Haas. • 1 Frasco (0,40 ml) de Activation Reagent (reactivo de activación). 30 mM de solución de cloruro de manganeso.
Armored RNA ist ein eingetragenes Warenzeichen von Ambion. Conservantes: ProClin 300 al 0,1% y ProClin 950 al 0,15%.
Abbott RealTime ist ein Warenzeichen von Abbott Laboratories. ProClin es una marca comercial registrada de Rohm and Haas.
Armored RNA es una marca comercial registrada de Ambion.
Abbott RealTime es una marca comercial de Abbott Laboratories.
(fr) Pour diagnostic in vitro. Abbott RealTime HCV est un dosage in vitro par RT-PCR (reverse transcription-polymerase chain reaction) pour la mesure
quantitative de l’acide ribonucléique du virus de l’hépatite C (ARN VHC) dans le sérum ou le plasma d’individus infectés par le VHC. Le dosage Abbott (it) Per uso diagnostico in vitro. Il dosaggio Abbott RealTime HCV è un dosaggio in vitro basato sul principio della RT-PCR (trascrizione inversa-reazione
a catena della polimerasi) per la determinazione quantitativa dell’acido ribonucleico del virus dell’epatite C (HCV RNA) in campioni di siero e plasma
RealTime HCV doit être utilisé comme aide dans le suivi des patients infectés par le VHC sous traitement antiviral. Le dosage Abbott RealTime HCV n’est
pas destiné au dépistage du VHC dans des échantillons sanguins, plasmatiques, sériques ou tissulaires de donneurs, ni à la confirmation du diagnostic umani prelevati da soggetti con infezione da HCV. Il dosaggio Abbott RealTime HCV è utilizzato come ausilio nella gestione dei pazienti con infezione da
d’une infection par le VHC. HCV e sottoposti a terapia antivirale. Il dosaggio Abbott RealTime HCV non deve essere utilizzato come test di screening del sangue, del plasma, del siero
Contenu : o dei tessuti donati per il virus dell’epatite C o come test diagnostico per confermare la presenza dell’infezione da HCV.
1. Contrôle interne Abbott RealTime HCV (4 flacons de 1,2 ml chacun). < 0,01 % d’Armored RNA® non Contenuto:
infectieux comprenant des séquences de contrôle interne dans du plasma humain négatif. Le plasma humain négatif a été testé et trouvé 1. Controllo interno Abbott RealTime HCV (4 provette, 1,2 ml per provetta). Armored RNA® non infettivo
non réactif pour l’AgHBs, l’ADN VHB, l’ARN VHC, l’ARN VIH et pour les anticorps anti-VIH-1/VIH-2 et anti-VHC. Conservateurs : <0,01% con sequenze di controllo interno in plasma umano negativo. Plasma umano negativo analizzato e risultato non reattivo per
ProClin® 300 à 0,1 % et ProClin 950 à 0,15 %. l’HBsAg, l’HBV DNA, l’HCV RNA, l’HIV RNA e per gli anticorpi anti‑HIV‑1/HIV-2 e anti-HCV. Conservanti: Proclin® 300 allo 0,1% e ProClin
2. Coffret-réactifs pour amplification Abbott RealTime HCV (4 boîtes de 24 tests chacune). Chaque 950 allo 0,15%.
coffret-réactifs contient : 2. Confezione dei reagenti per l’amplificazione Abbott RealTime HCV (4 confezioni, 24 test/
• 1 flacon (0,141 ml) d’enzyme rTth polymérase thermostable (2,9 à 3,5 unités/µl) dans une solution tampon. confezione). Ciascuna confezione dei reagenti contiene:
• 1 flacon (1,10 ml) de réactif d’oligonucléotides de VHC. < 0,1 % d’oligonucléotides synthétiques (4 amorces et 2 sondes) et • 1 flacone (0,141 ml) di enzima termostabile polimerasi rTth (da 2,9 a 3,5 unità/µl) in soluzione tamponata.
< 0,3 % de dNTPs dans une solution tampon contenant un fluorochrome de référence. Conservateurs : ProClin 300 à 0,1 % et • 1 flacone (1,10 ml) di reagente di oligonucleotidi dell’HVC. Oligonucleotidi sintetici <0,1% (4 primer e 2 sonde), e dNTPs <0,3% in
ProClin 950 à 0,15 %. una soluzione tamponata con un colorante di riferimento. Conservanti: ProClin 300 allo 0,1% e ProClin 950 allo 0,15%.
• 1 flacon (0,40 ml) de réactif d’activation. 30 mM de solution de chlorure de manganèse. Conservateurs : ProClin 300 à 0,1 % et • 1 flacone (0,40 ml) di reagente di attivazione. 30 mM di soluzione di cloruro di manganese. Conservanti: ProClin 300 allo 0,1% e
ProClin 950 à 0,15 %. ProClin 950 allo 0,15%.
ProClin est une marque déposée de Rohm and Haas. ProClin è un marchio commerciale registrato di Rohm and Haas.
Armored RNA est une marque déposée d’Ambion. Armored RNA è un marchio commerciale registrato di Ambion.
Abbott RealTime est une marque commerciale d’Abbott Laboratories. Abbott RealTime è un marchio commerciale di Abbott Laboratories.
PQDx 0450-027-00 WHO PQ Public Report November 2021, version 3.0

1.1.1 Amplification reagent label

1.2 Abbott RealTime HCV Control Kit box label (4J86-80)

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5/29/2020 10:56:21 AM 51-602120R5.indd 1
EE.UU. / Product of USA / Prodotto in USA / Fabricado nos EUA
Product of USA / Produkt aus USA / Produit des Etats-Unis / Producto de Abbott RealTime é uma marca comercial de Abbott Laboratories.
Armored RNA é uma marca comercial de Ambion.
ProClin é uma marca comercial registada de Rohm and Haas.
e anti-HCV. Conservantes: 0,1% de ProClin 300 e 0,15% de ProClin 950.
humano negativo. Plasma humano negativo testado e considerado não-reactivo para HBsAg, ADN do HBV, ARN do HCV, ARN do HIV, anticorpos anti-HIV-1/HIV-2
3. Abbott RealTime HCV High Positive Control (8 frascos, 1,3 ml por frasco). ARN protegido não-infeccioso com sequências de HCV em plasma
anticorpos anti-HIV-1/HIV-2 e anti-HCV. Conservantes: 0,1% de ProClin 300 e 0,15% de ProClin 950.
em plasma humano negativo. Plasma humano negativo testado e considerado não-reactivo para HBsAg, ADN do HBV, ARN do HCV, ARN do HIV,
2. Abbott RealTime HCV Low Positive Control (8 frascos, 1,3 ml por frasco). Armored RNA® (ARN protegido) não-infeccioso com sequências de HCV
anticorpos anti-HIV-1/HIV-2 e anti-HCV. Conservantes: 0,1% de ProClin® 300 e 0,15% de ProClin 950.
ADN do HBV, ARN do HCV, ARN do HIV,
1. Abbott RealTime HCV Negative Control (8 frascos, 1,8 ml por frasco). Plasma humano negativo testado e considerado não-reactivo para HBsAg,
Conteúdo:
quantificação do ARN do vírus da hepatite C (HCV) em soro e plasma humanos de indivíduos infectados pelo HCV.
(pt) Para utilização in vitro. Os Abbott RealTime HCV Controls são utilizados para estabelecer a validade do ensaio Abbott RealTime HCV quando utilizado para a
HCV
Control Kit Abbott RealTime
Consultar as instruções de utilização.
ATENÇÃO: manusear os materiais de origem humana como potencialmente infecciosos.
i materiali di origine umana come potenzialmente infettivi. Consultare le istruzioni per l’uso. /
como potencialmente infecciosos. Consulte las instrucciones de uso. / ATTENZIONE: Trattare
Consulter les instructions d’utilisation. / ATENCIÓN: maneje los productos de origen humano
: Manipuler les produits d’origine humaine comme s’ils étaient potentiellement infectieux.
entsprechenden Vorsicht gehandhabt werden. Gebrauchsanweisung beachten. / ATTENTION
for use. / ACHTUNG: Humanmaterial gilt als potentiell infektiös und muss mit der
CAUTION: Handle human sourced materials as potentially infectious. Consult instructions
Abbott RealTime is a trademark of Abbott Laboratories.
51-602120/R5 Armored RNA is a trademark of Ambion.
ProClin is a registered trademark of Rohm and Haas.
Preservatives: 0.1% ProClin 300 and 0.15% ProClin 950.
found to be nonreactive for HBsAg, HBV DNA, HCV RNA, HIV RNA, anti-HIV-1/HIV-2, and anti-HCV.
Armored RNA with HCV sequences in negative human plasma. Negative human plasma tested and
1434
3. Abbott RealTime HCV High Positive Control (8 vials, 1.3 mL per vial). Noninfectious
Preservatives: 0.1% ProClin 300 and 0.15% ProClin 950.
found to be nonreactive for HBsAg, HBV DNA, HCV RNA, HIV RNA, anti-HIV-1/HIV-2, and anti-HCV.
Armored RNA® with HCV sequences in negative human plasma. Negative human plasma tested and
2. Abbott RealTime HCV Low Positive Control (8 vials, 1.3 mL per vial). Noninfectious
anti-HIV-1/HIV-2, and anti-HCV. Preservatives: 0.1% ProClin® 300 and 0.15% ProClin 950.
plasma tested and found to be nonreactive for HBsAg, HBV DNA, HCV RNA, HIV RNA,
1. Abbott RealTime HCV Negative Control (8 vials, 1.8 mL per vial). Negative human
P333+P313, P362+P364, P501 Contents:
P261, P280, P272, P302+P352,
and plasma from HCV infected individuals.
H317
Abbott RealTime HCV assay when used for the quantitation of hepatitis C virus (HCV) RNA in human serum
4J86-80 (en) For In Vitro Diagnostic Use. The Abbott RealTime HCV Controls are used to establish run validity of the
HCV
Control Kit Abbott RealTime
(de) In-vitro-Diagnostikum. Die Abbott RealTime HCV Kontrollen dienen zur Herstellung der Testgültigkeit des Abbott RealTime HCV (es) Para uso en diagnóstico in vitro. Los controles Abbott RealTime HCV se usan para establecer la validez del procesamiento del
Assays bei der quantitativen Bestimmung von Hepatitis-C-viraler Ribonukleinsäure (HCV RNA) in Humanserum und -plasma von ensayo Abbott RealTime HCV en la determinación cuantitativa del RNA del virus de la hepatitis C (VHC) en plasma y suero humano de
HCV-infizierten Personen. individuos infectados con el VHC.
Inhalt: Contenido:
1. Abbott RealTime HCV Negative Kontrolle (8 Fläschchen, 1,8 ml pro Fläschchen). Negatives Humanplasma wurde 1. Control negativo Abbott RealTime HCV (8 tubos de 1,8 ml cada uno). El plasma humano negativo se ha analizado
getestet und war nicht reaktiv für HBsAg, HBV DNA, HCV RNA, HIV RNA, anti-HIV‑1/HIV‑2 und anti-HCV. Konservierungsmittel: y no se ha encontrado reactividad para el HBsAg, ni para el DNA del VHB, ni el RNA del VHC, ni el RNA del VIH, ni reactividad de
0,1 % ProClin® 300 und 0,15 % ProClin 950. anticuerpos anti-VIH-1/VIH-2 ni anti-VHC. Conservantes: ProClin® 300 al 0,1% y ProClin 950 al 0,15%.
2. Abbott RealTime HCV Niedrig Positive Kontrolle (8 Fläschchen, 1,3 ml pro Fläschchen). Nicht infektiöse Armored 2. Control positivo bajo Abbott RealTime HCV (8 tubos de 1,3 ml cada uno). Armored RNA® no infeccioso con
RNA® mit HCV-Sequenzen in negativem Humanplasma. Negatives Humanplasma wurde getestet und war nicht reaktiv für HBsAg, secuencias de VHC en plasma humano negativo. El plasma humano negativo se ha analizado y no se ha encontrado reactividad
HBV DNA, HCV RNA, HIV RNA, anti-HIV-1/HIV-2 und anti-HCV. Konservierungsmittel: 0,1 % ProClin 300 und 0,15 % ProClin 950. para el HBsAg, ni para el DNA del VHB, ni el RNA del VHC, ni el RNA del VIH, ni reactividad de anticuerpos anti-VIH-1/VIH-2 ni
3. Abbott RealTime HCV Hoch Positive Kontrolle (8 Fläschchen, 1,3 ml pro Fläschchen). Nicht infektiöse Armored anti-VHC. Conservantes: ProClin® 300 al 0,1% y ProClin 950 al 0,15%.
RNA mit HCV-Sequenzen in negativem Humanplasma. Negatives Humanplasma wurde getestet und war nicht reaktiv für HBsAg, 3. Control positivo alto Abbott RealTime HCV (8 tubos de 1,3 ml cada uno). Armored RNA no infeccioso con
HBV DNA, HCV RNA, HIV RNA, anti-HIV-1/HIV-2 und anti-HCV. Konservierungsmittel: 0,1 % ProClin 300 und 0,15 % ProClin 950. secuencias de VHC en plasma humano negativo. El plasma humano negativo se ha analizado y no se ha encontrado reactividad
ProClin ist ein eingetragenes Warenzeichen von Rohm and Haas. para el HBsAg, ni para el DNA del VHB, ni el RNA del VHC, ni el RNA del VIH, ni reactividad de anticuerpos anti-VIH-1/VIH-2 ni
Armored RNA ist ein Warenzeichen von Ambion. anti-VHC. Conservantes: ProClin® 300 al 0,1% y ProClin 950 al 0,15%.
Abbott RealTime ist ein Warenzeichen von Abbott Laboratories. ProClin es una marca comercial registrada de Rohm and Haas.
(fr) Pour diagnostic in vitro. Les contrôles Abbott RealTime HCV sont utilisés pour établir la validité des analyses effectuées avec Armored RNA es una marca comercial de Ambion.
le dosage Abbott RealTime HCV lors de la mesure quantitative de l’ARN du virus de l’hépatite C (VHC) dans le sérum ou le plasma Abbott RealTime es una marca comercial de Abbott Laboratories.
d’individus infectés par le VHC. (it) Per uso diagnostico in vitro. I controlli Abbott RealTime HCV vengono utilizzati per stabilire la validità dell’analisi con il dosaggio
Contenu : Abbott RealTime HCV per la determinazione quantitativa dell’RNA del virus dell’epatite C (HCV) in campioni di siero e plasma umani
1. Contrôle négatif Abbott RealTime HCV (8 flacons de 1,8 ml chacun). Le plasma humain négatif a été testé et prelevati da soggetti con infezione da HCV.
trouvé non réactif pour l’AgHBs, l’ADN VHB, l’ARN VHC, l’ARN VIH et pour les anticorps anti-VIH-1/VIH-2 et anti-VHC. Conservateurs Contenuto:
: ProClin® 300 à 0,1 % et ProClin 950 à 0,15 %. 1. . Controllo negativo Abbott RealTime HCV (8 provette, 1,8 ml per provetta). Plasma umano negativo analizzato e
2. Contrôle positif bas Abbott RealTime HCV (8 flacons de 1,3 ml chacun). Armored RNA® non infectieux risultato non reattivo per l’HBsAg, l’HBV DNA, l’HCV RNA, l’HIV RNA e per gli anticorpi anti-HIV‑1/HIV‑2 e anti-HCV. Conservanti:
comprenant des séquences VHC dans du plasma humain négatif. Le plasma humain négatif a été testé et trouvé non réactif pour ProClin® 300 allo 0,1% e ProClin 950 allo 0,15% .
l’AgHBs, l’ADN VHB, l’ARN VHC, l’ARN VIH et pour les anticorps anti-VIH-1/VIH-2 et anti‑VHC. Conservateurs : ProClin® 300 à 0,1 % 2. . Controllo positivo basso Abbott RealTime HCV (8 provette, 1,3 ml per provetta). Armored RNA® non infettivo con
et ProClin 950 à 0,15 %. sequenze di HCV in plasma umano negativo. Plasma umano negativo analizzato e risultato non reattivo per l’HBsAg, l’HBV DNA,
3. Contrôle positif haut Abbott RealTime HCV (8 flacons de 1,3 ml chacun). Armored RNA non infectieux comprenant l’HCV RNA, l’HIV RNA e per gli anticorpi anti-HIV-1/HIV-2 e anti-HCV. Conservanti: ProClin 300 allo 0,1% e ProClin 950 allo 0,15%.
des séquences VHC dans du plasma humain négatif. Le plasma humain négatif a été testé et trouvé non réactif pour l’AgHBs, 3. Controllo positivo alto Abbott RealTime HCV (8 provette, 1,3 ml per provetta). Armored RNA non infettivo con
l’ADN VHB, l’ARN VHC, l’ARN VIH et pour les anticorps anti-VIH-1/VIH-2 et anti‑VHC. Conservateurs : ProClin® 300 à 0,1 % et sequenze di HCV in plasma umano negativo. Plasma umano negativo analizzato e risultato non reattivo per l’HBsAg, l’HBV DNA,
ProClin 950 à 0,15 %. l’HCV RNA, l’HIV RNA e per gli anticorpi anti-HIV-1/HIV-2 e anti-HCV. Conservanti: ProClin 300 allo 0,1% e ProClin 950 allo 0,15%.
ProClin est une marque déposée de Rohm and Haas. ProClin è un marchio commerciale registrato di Rohm and Haas.
Armored RNA est une marque commerciale d’Ambion. Armored RNA è un marchio commerciale di Ambion.
Abbott RealTime est une marque commerciale d’Abbott Laboratories. Abbott RealTime è un marchio commerciale di Abbott Laboratories.
PQDx 0450-027-00 WHO PQ Public Report November 2021, version 3.0

1.2.1 Abbott RealTime HCV Negative Control vial Label (4J86Z)

1.2.2 Abbott RealTime HCV Low Positive Control vial Label (4J86W)

Page 12 of 32
PQDx 0450-027-00 WHO PQ Public Report November 2021, version 3.0

1.2.3 Abbott RealTime HCV High Positive Control vial Label (4J86X)

1.3 Abbott RealTime HCV Calibrator Kit box label (4J86-70)

Page 13 of 32
5/29/2020 10:52:55 AM 51-602115R5.indd 1
www.abbottmolecular.com Des Plaines, IL 60018 USA
1300 East Touhy Avenue
Abbott Molecular Inc.
Producto de EE.UU. / Prodotto degli USA / Fabricado nos EUA
Product of USA / Produkt aus USA / Produit aux Etats-Unis /
Abbott RealTime é uma marca comercial de Abbott Laboratories.
Armored RNA é uma marca comercial registada de Ambion.
ProClin é uma marca comercial registada de Rohm and Haas.
HIV, anticorpos anti‑HIV‑1/HIV-2 e anti-HCV. Conservantes: 0,1% de ProClin 300 e 0,15% de ProClin 950.
plasma humano negativo. Plasma humano negativo testado e considerado não-reactivo para HBsAg, ADN do HBV, ARN do HCV, ARN do
2. Abbott RealTime HCV Calibrator B (12 frascos, 1,3 ml por frasco). ARN protegido não-infeccioso com sequências de HCV em
ProClin 950.
ARN do HCV, ARN do HIV, anticorpos anti‑HIV‑1/HIV-2 e anti-HCV. Conservantes: 0,1% de ProClin® 300 e 0,15% de
sequências de HCV em plasma humano negativo. Plasma humano negativo testado e considerado não-reactivo para HBsAg, ADN do HBV,
1. Abbott RealTime HCV Calibrator A (12 frascos, 1,3 ml por frasco). Armored RNA® (ARN protegido) não-infeccioso com
Conteúdo:
determinação quantitativa do ARN do vírus da hepatite C (HCV) em soro e plasma humanos de indivíduos infectados pelo HCV.
(pt) Para utilização in vitro. Os Abbott RealTime HCV Calibrators destinam-se à calibração do ensaio Abbott RealTime HCV, quando utilizado para a
HCV
Calibrator Kit Abbott RealTime
como potencialmente infecciosos. Consultar as instruções de utilização.
Consultare le istruzioni per l’uso. / ATENÇÃO: manusear os materiais de origem humana
uso. / ATTENZIONE: Trattare i materiali di origine umana come potenzialmente infettivi.
de origen humano como potencialmente infecciosos. Consulte las instrucciones de
infectieux. Consulter les instructions d’utilisation. / ATENCIÓN: maneje los productos
ATTENTION : Manipuler les produits d’origine humaine comme s’ils étaient potentiellement
mit der entsprechenden Vorsicht gehandhabt werden. Gebrauchsanweisung beachten. /
instructions for use. / ACHTUNG: Humanmaterial gilt als potentiell infektiös und muss
CAUTION: Handle human sourced materials as potentially infectious. Consult
Abbott RealTime is a trademark of Abbott Laboratories.
51-602115/R5
Armored RNA is a registered trademark of Ambion.
ProClin is a registered trademark of Rohm and Haas.
ProClin 950.
RNA, anti‑HIV‑1/HIV-2, and anti-HCV. Preservatives: 0.1% ProClin 300 and 0.15%
plasma tested and found to be nonreactive for HBsAg, HBV DNA, HCV RNA, HIV
Armored RNA with HCV sequences in negative human plasma. Negative human
2. Abbott RealTime HCV Calibrator B (12 vials, 1.3 mL per vial). Noninfectious
ProClin 950.
RNA, anti‑HIV‑1/HIV-2, and anti-HCV. Preservatives: 0.1% ProClin® 300 and 0.15%
1434
plasma tested and found to be nonreactive for HBsAg, HBV DNA, HCV RNA, HIV
Armored RNA® with HCV sequences in negative human plasma. Negative human
1. Abbott RealTime HCV Calibrator A (12 vials, 1.3 mL per vial). Noninfectious
P333+P313, P362+P364, P501 Contents:
P261, P280, P272, P302+P352, virus (HCV) RNA in human serum and plasma from HCV infected individuals.
H317 the Abbott RealTime HCV assay when used for the quantitative determination of hepatitis C
4J86-70 (en) For In Vitro Diagnostic Use. The Abbott RealTime HCV Calibrators are for calibration of
HCV
Calibrator Kit Abbott RealTime
(de) In-vitro-Diagnostikum. Die Abbott RealTime HCV Kalibratoren dienen zur Kalibrierung des Abbott RealTime HCV (es) Para uso en diagnóstico in vitro. Los calibradores Abbott RealTime HCV se usan para la calibración del ensayo Abbott
Assays bei der quantitativen Bestimmung von Hepatitis-C-viraler Ribonukleinsäure (HCV RNA) in Humanserum und RealTime HCV en la determinación cuantitativa del RNA del virus de la hepatitis C (VHC) en suero y plasma humano de
-plasma von HCV-infizierten Personen. individuos infectados con el VHC.
Inhalt: Contenido:
1. Abbott RealTime HCV Kalibrator A (12 Fläschchen, 1,3 ml pro Fläschchen). Nicht infektiöse 1. Calibrador A Abbott RealTime HCV (12 tubos de 1,3 ml cada uno). Armored RNA® no infeccioso
Armored RNA® mit HCV-Sequenzen in negativem Humanplasma. Negatives Humanplasma wurde con secuencias de VHC en plasma humano negativo. El plasma humano negativo se ha analizado y no se
getestet und war nicht reaktiv für HBsAg, HBV DNA, HCV RNA, HIV RNA, anti-HIV-1/HIV-2 und anti-HCV. ha encontrado reactividad para el HBsAg, ni para el DNA del VHB, ni el RNA del VHC, ni el RNA del VIH, ni
Konservierungsmittel: 0,1 % ProClin® 300 und 0,15 % ProClin 950. reactividad de anticuerpos anti-VIH-1/VIH-2 ni anti-VHC. Conservantes: ProClin® 300 al 0,1% y ProClin 950 al
2. Abbott RealTime HCV Kalibrator B (12 Fläschchen, 1,3 ml pro Fläschchen). Nicht infektiöse 0,15%.
Armored RNA mit HCV-Sequenzen in negativem Humanplasma. Negatives Humanplasma wurde getestet 2. Calibrador B Abbott RealTime HCV (12 tubos de 1,3 ml cada uno). Armored RNA no infeccioso
und war nicht reaktiv für HBsAg, HBV DNA, HCV RNA, HIV RNA, anti‑HIV‑1/HIV-2 und anti-HCV. con secuencias de VHC en plasma humano negativo. El plasma humano negativo se ha analizado y no se
Konservierungsmittel: 0,1 % ProClin 300 und 0,15 % ProClin 950. ha encontrado reactividad para el HBsAg, ni para el DNA del VHB, ni el RNA del VHC ni el RNA del VIH, ni
ProClin ist ein eingetragenes Warenzeichen von Rohm and Haas. reactividad de anticuerpos anti‑VIH‑1/VIH-2 ni anti-VHC. Conservantes: ProClin® 300 al 0,1% y ProClin 950 al
Armored RNA ist ein eingetragenes Warenzeichen von Ambion. 0,15%.
Abbott RealTime ist ein Warenzeichen von Abbott Laboratories. ProClin es una marca comercial registrada de Rohm and Haas.
Armored RNA es una marca comercial registrada de Ambion.
(fr) Pour diagnostic in vitro. Les calibrateurs Abbott RealTime HCV sont utilisés pour la calibration du dosage Abbott Abbott RealTime es una marca comercial de Abbott Laboratories.
RealTime HCV lors de la mesure quantitative de l’ARN du virus de l’hépatite C (HCV) dans du sérum ou du plasma
d’individus infectés par le VHC.
Contenu : (it) Per uso diagnostico in vitro. I calibratori Abbott RealTime HCV vengono utilizzati per la calibrazione del dosaggio Abbott
1. Calibrateur A Abbott RealTime HCV (12 flacons de 1,3 ml chacun). Armored RNA® non infectieux RealTime HCV per la determinazione quantitativa dell’RNA del virus dell’epatite C (HCV) in campioni di siero e plasma
comprenant des séquences de VHC dans du plasma humain négatif. Le plasma humain négatif a été testé et umani prelevati da soggetti con infezione da HCV.
trouvé non réactif pour l’AgHBs, l’ADN VHB, l’ARN VHC, l’ARN VIH et pour les anticorps anti-VIH-1/VIH-2 et Contenuto:
anti-VHC. Conservateurs : ProClin® 300 à 0,1 % et ProClin 950 à 0,15 %. 1. Calibratore A Abbott RealTime HCV (12 provette, 1,3 ml per provetta). Armored RNA® non infettivo
2. Calibrateur B Abbott RealTime HCV (12 flacons de 1,3 ml chacun). Armored RNA non infectieux con sequenze di HCV in plasma umano negativo. Plasma umano negativo analizzato e risultato non reattivo
comprenant des séquences de VHC dans du plasma humain négatif. Le plasma humain négatif a été testé et per l’HBsAg, l’HBV DNA, l’HCV RNA, l’HIV RNA e per gli anticorpi anti-HIV-1/HIV-2 e anti‑HCV. Conservanti:
trouvé non réactif pour l’AgHBs, l’ADN VHB, l’ARN VHC, l’ARN VIH et pour les anticorps anti-VIH-1/VIH-2 et ProClin® 300 allo 0,1% e ProClin 950 allo 0,15% .
anti-VHC. Conservateurs : ProClin 300 à 0,1 % et ProClin 950 à 0,15 %. 2. Calibratore B Abbott RealTime HCV (12 provette, 1,3 ml per provetta). Armored RNA non infettivo
ProClin est une marque déposée de Rohm and Haas. con sequenze di HCV in plasma umano negativo. Plasma umano negativo analizzato e risultato non reattivo per
Armored RNA est une marque déposée d’Ambion. l’HBsAg, l’HBV DNA, l’HCV RNA, l’HIV RNA e per gli anticorpi anti-HIV-1/HIV-2 e anti‑HCV. Conservanti: ProClin
Abbott RealTime est une marque commerciale d’Abbott Laboratories. 300 allo 0,1% e ProClin 950 allo 0,15%.
ProClin è un marchio commerciale registrato di Rohm and Haas.
Armored RNA è un marchio commerciale registrato di Ambion.
Abbott RealTime è un marchio commerciale di Abbott Laboratories.
PQDx 0450-027-00 WHO PQ Public Report November 2021, version 3.0

1.3.1 Abbott RealTime HCV CAL A vial label (4J86A)

1.3.2 Abbott RealTime HCV CAL B vial label (4J86B)

Page 15 of 32
PQDx 0450-027-00 WHO PQ Public Report November 2021, version 3.0

2. Instructions for use 1

1
English version of the IFU was the one that was assessed by WHO. It is the responsibility of the
manufacturer to ensure correct translation into other languages.

Page 16 of 32
 Abbott RealTime IN VITRO TEST
en
4J86-70

HCV Calibrators 51-602116/R6

NOTE: Changes Highlighted
Standardization
Key to Symbols Used
Abbott manufactures internal reference standards for the Abbott
List Number RealTime HCV assay. These internal standards are referenced to the
Second WHO International Standard for Hepatitis C Virus RNA (NIBSC
In Vitro Diagnostic Medical Device Code 96/798)1 at each concentration level. The Abbott RealTime HCV
calibrators are manufactured against these internal standards.
Lot Number Precautions
• In Vitro Diagnostic Medical Device
Expiration Date • For In Vitro Diagnostic Use Only
• Do not use beyond expiration date.
Calibrator (A - B)
CAUTION: This preparation contains human sourced and/or
Store at ≤ -10°C potentially infectious components. Components sourced from human
blood have been tested and found to be nonreactive by FDA-licensed
tests for antibody to HCV, antibody to HIV-1, antibody to HIV-2, and
Consult instructions for use HBsAg. The material is also tested and found to be negative by FDA-
licensed PCR methods for HIV-1 RNA and HCV RNA. No known test
method can offer complete assurance that products derived from human
sources or inactivated microorganisms will not transmit infection. These
Warning reagents and human specimens should be handled as if infectious
using safe laboratory procedures, such as those outlined in Biosafety
CAUTION: Handle human sourced materials as in Microbiological and Biomedical Laboratories,2 OSHA Standards on
potentially infectious. Consult instructions for use. Bloodborne Pathogens,3 CLSI Document M29-A3,4 and other appropriate
(Infection Risk) biosafety practices.5 Therefore all human sourced materials should be
considered infectious.
Authorized Representative These precautions include, but are not limited to, the following:
• Wear gloves when handling specimens or reagents.
Manufacturer • Do not pipette by mouth.
• Do not eat, drink, smoke, apply cosmetics, or handle contact lenses
in areas where these materials are handled.
Notice to User • Clean and disinfect spills of specimens by including the use of a
If a serious incident occurs in relation to this device, the incident should tuberculocidal disinfectant such as 1.0% sodium hypochlorite or
be reported to the manufacturer and to the appropriate competent other suitable disinfectant.2
authority of the member state in which the user and/or the patient is
established. To report to the manufacturer, see the contact information • Decontaminate and dispose of all potentially infectious materials in
provided in the Customer service section or Technical assistance accordance with local, state and federal regulations.5
section of these instructions. Components of the Abbott RealTime HCV Calibrator Kit
(List No. 4J86-70) contain the following components:
Intended Use • 2-Methyl-2H-isothiazol-3-one
The Abbott RealTime HCV Calibrators are for calibration of the Abbott
• Reaction mass of: 5-chloro-2-methyl-4-isothiazolin-3-one
RealTime HCV assay when used for the quantitative determination of
(EC no. 247-500-7) and 2-methyl-2H-isothiazol-3-one
Hepatitis C Virus (HCV) RNA in human serum and plasma from HCV
(EC no. 220-239-6)(3:1)
infected individuals.
• Reaction mass of: 5-chloro-2-methyl-4-isothiazolin-3-one
Intended User (EC no. 247-500-7) and 2-methyl-4-isothiazolin-3-one
The intended users for the Abbott RealTime HCV Calibrators are (EC no. 220-239-6)(3:1)
laboratory and healthcare professionals. The following warnings apply:
Contents Warning
1. Abbott RealTime HCV Calibrator A (List No. 4J86A) H317 May cause an allergic skin reaction.
(12 vials, 1.3 mL per vial). Noninfectious Armored RNA® with HCV
sequences in negative human plasma. Negative human plasma P261 Avoid breathing mist / vapours / spray.
tested and found to be nonreactive for HBsAg, HBV DNA, HIV RNA, P280 Wear protective gloves / protective clothing /
HCV RNA, anti‑HIV‑1/HIV‑2, and anti-HCV. Preservatives: 0.1% eye protection.
ProClin® 300 and 0.15% ProClin 950. P272 Contaminated work clothing should not be
2. Abbott RealTime HCV Calibrator B (List No. 4J86B) allowed out of the workplace.
(12 vials, 1.3 mL per vial). Noninfectious Armored RNA with HCV P302+P352 IF ON SKIN: Wash with plenty of water.
sequences in negative human plasma. Negative human plasma P333+P313 If skin irritation or rash occurs: Get medical
tested and found to be nonreactive for HBsAg, HBV DNA, HIV RNA, advice / attention.
HCV RNA, anti‑HIV‑1/HIV‑2, and anti-HCV. Preservatives: 0.1%
ProClin 300 and 0.15% ProClin 950. P362+P364 Take off contaminated clothing and wash it
before reuse.
• Calibrator concentrations are specified in each Abbott RealTime P501 Dispose of contents / container in accordance
HCV Calibrator Kit Card. with local regulations.
• The Abbott RealTime HCV Calibrator Kit must only be used with the
Abbott RealTime HCV assay (List No. 4J86-90).

WHO Change Notification

Volume 1
Page 135 of 156
 Consult instructions for use

Store at ≤ -10°C

Shipping Conditions
Ship on dry ice.
BIBLIOGRAPHY
1. Saldanha J, Lelie N, Heath A. Establishment of the first international
standard for nucleic acid amplification technology (NAT) assays for
HCV RNA. Vox Sang. 1999;76:149-58.
2. US Department of Health and Human Services. Biosafety in
Microbiological and Biomedical Laboratories. 5th ed. Washington,
DC: US Government Printing Office; December 2009.
[Also available online. Type> www.cdc.gov, search>BMBL5>look up
sections III and IV.]
3. US Department of Labor, Occupational Safety and Health
Administration. 29 CFR Part 1910.1030. Bloodborne Pathogens.
4. Clinical and Laboratory Standards Institute. Protection of Laboratory
Workers from Occupationally Acquired Infections: Approved
Guideline—Third Edition. CLSI Document M29-A3. Wayne, PA:
Clinical and Laboratory Standards Institute; 2005.
5. World Health Organization. Laboratory Biosafety Manual. 3rd ed.
Geneva, Switzerland: World Health Organization; 2004.
Technical Assistance
For technical assistance, call Abbott Molecular Technical Services at
1-800-553-7042 (within the US) or +49-6122-580 (outside the US), or
visit the Abbott Molecular website at www.molecular.abbott/portal.
Armored RNA is a registered trademark of Ambion.
ProClin is a registered trademark of Rohm and Haas.
Abbott RealTime is a trademark of Abbott Laboratories.
The Abbott RealTime HCV Calibrators Kit is imported into the European
Union by Abbott Diagnostics GmbH, located at Max-Planck-Ring 2,
65205 Wiesbaden, Germany.

1434

www.abbottmolecular.com
©2005, 2020 Abbott. All Rights Reserved.
June 2020

2
 Abbott RealTime en
IN VITRO TEST
4J86-80

HCV Controls 51-602121/R6

NOTE: Changes Highlighted
• Control concentrations are specified in each Abbott RealTime HCV
Key to Symbols Used Control Kit Card.
List Number • The Abbott RealTime HCV Control Kit must only be used with the
Abbott RealTime HCV assay (List No. 4J86-90).
In Vitro Diagnostic Medical Device Precautions
• In Vitro Diagnostic Medical Device
Lot Number
• For In Vitro Diagnostic Use Only
Negative Control • Do not use beyond expiration date.

Positive Control Low

CAUTION: This preparation contains human sourced and/or
Positive Control High potentially infectious components. Components sourced from human
blood have been tested and found to be nonreactive by FDA-licensed
tests for antibody to HCV, antibody to HIV-1, antibody to HIV-2, and
Store at ≤ -10°C HBsAg. The material is also tested and found to be negative by FDA-
licensed PCR methods for HIV-1 RNA and HCV RNA. No known test
method can offer complete assurance that products derived from human
Warning sources or inactivated microorganisms will not transmit infection. These
reagents and human specimens should be handled as if infectious
using safe laboratory procedures, such as those outlined in Biosafety
in Microbiological and Biomedical Laboratories,1 OSHA Standards on
Consult instructions for use Bloodborne Pathogens,2 CLSI Document M29-A3,3 and other appropriate
biosafety practices.4 Therefore all human sourced materials should be
CAUTION: Handle human sourced materials as considered infectious.
potentially infectious. Consult instructions for use. These precautions include, but are not limited to, the following:
(Infection Risk) • Wear gloves when handling specimens or reagents.
Authorized Representative • Do not pipette by mouth.
• Do not eat, drink, smoke, apply cosmetics, or handle contact lenses
in areas where these materials are handled.
Manufacturer • Clean and disinfect spills of specimens by including the use of a
tuberculocidal disinfectant such as 1.0% sodium hypochlorite or
Notice to User other suitable disinfectant.1
If a serious incident occurs in relation to this device, the incident should • Decontaminate and dispose of all potentially infectious materials in
be reported to the manufacturer and to the appropriate competent accordance with local, state and federal regulations.4
authority of the member state in which the user and/or the patient is Components of the Abbott RealTime HCV Control Kit (List No. 4J86-80)
established. To report to the manufacturer, see the contact information contain the following components:
provided in the Customer service section or Technical assistance • 2-Methyl-2H-isothiazol-3-one
section of these instructions. • Reaction mass of: 5-chloro-2-methyl-4-isothiazolin-3-one
Intended Use (EC no. 247-500-7) and 2-methyl-2H-isothiazol-3-one
The Abbott RealTime HCV Controls are used to establish run validity (EC no. 220-239-6)(3:1)
of the Abbott RealTime HCV assay when used for the quantitation of • Reaction mass of: 5-chloro-2-methyl-4-isothiazolin-3-one
Hepatitis C Virus (HCV) RNA in human serum and plasma from HCV (EC no. 247-500-7) and 2-methyl-4-isothiazolin-3-one
infected individuals. (EC no. 220-239-6)(3:1)
Intended User The following warnings apply:
The intended users for the Abbott RealTime HCV Controls are laboratory Warning
and healthcare professionals. H317 May cause an allergic skin reaction.
Contents P261 Avoid breathing mist / vapours / spray.
1. Abbott RealTime HCV Negative Control P280 Wear protective gloves / protective clothing /
(List No. 4J86Z) (8 vials, 1.8 mL per vial). Negative human plasma. eye protection.
Negative human plasma tested and found to be nonreactive for P272 Contaminated work clothing should not be
HBsAg, HBV DNA, HIV RNA, HCV RNA, anti-HIV-1/HIV-2, and allowed out of the workplace.
anti‑HCV. Preservatives: 0.1% ProClin® 300 and 0.15% ProClin 950.
P302+P352 IF ON SKIN: Wash with plenty of water.
2. Abbott RealTime HCV Low Positive Control
(List No. 4J86W) (8 vials, 1.3 mL per vial). Noninfectious Armored P333+P313 If skin irritation or rash occurs: Get medical
®
RNA with HCV sequences in negative human plasma. Negative advice / attention.
human plasma tested and found to be nonreactive for HBsAg, P362+P364 Take off contaminated clothing and wash it
HBV DNA, HIV RNA, HCV RNA, anti-HIV-1/HIV-2, and anti‑HCV. before reuse.
Preservatives: 0.1% ProClin 300 and 0.15% ProClin 950. P501 Dispose of contents / container in accordance
3. Abbott RealTime HCV High Positive Control with local regulations.
(List No. 4J86X) (8 vials, 1.3 mL per vial). Noninfectious Armored
RNA with HCV sequences in negative human plasma. Negative
human plasma tested and found to be nonreactive for HBsAg,
HBV DNA, HIV RNA, HCV RNA, anti-HIV-1/HIV-2, and anti‑HCV.
Preservatives: 0.1% ProClin 300 and 0.15% ProClin 950.
1
 Consult instructions for use

Store at ≤ -10°C

Shipping Conditions
Ship on dry ice.
BIBLIOGRAPHY
1. US Department of Health and Human Services. Biosafety in
Microbiological and Biomedical Laboratories. 5th ed. Washington,
DC: US Government Printing Office; December 2009.
[Also available online. Type> www.cdc.gov, search>BMBL5>look up
sections III and IV.]
2. US Department of Labor, Occupational Safety and Health
Administration. 29 CFR Part 1910.1030. Bloodborne Pathogens.
3. Clinical and Laboratory Standards Institute. Protection of Laboratory
Workers from Occupationally Acquired Infections: Approved
Guideline—Third Edition. CLSI Document M29-A3. Wayne, PA:
Clinical and Laboratory Standards Institute; 2005.
4. World Health Organization. Laboratory Biosafety Manual. 3rd ed.
Geneva, Switzerland: World Health Organization; 2004.
Technical Assistance
For technical assistance, call Abbott Molecular Technical Services at
1-800-553-7042 (within the US) or +49-6122-580 (outside the US), or
visit the Abbott Molecular website at www.molecular.abbott/portal.
Armored RNA is a registered trademark of Ambion.
ProClin is a registered trademark of Rohm and Haas.
Abbott RealTime is a trademark of Abbott Laboratories.
The Abbott RealTime HCV Controls Kit is imported into the European
Union by Abbott Diagnostics GmbH, located at Max-Planck-Ring 2,
65205 Wiesbaden, Germany.

1434

www.abbottmolecular.com
© 2005, 2020 Abbott. All Rights Reserved.
June 2020

2
 Abbott RealTime
HCV en
4J86

4J86 51-602124/R12
51-602124/R12
NOTE: Changes Highlighted
CUSTOMER SERVICE
Key to Symbols Used
INTERNATIONAL: CALL YOUR ABBOTT
Reference Number REPRESENTATIVE
This package insert must be read carefully prior to use. Package insert
Lot Number
instructions must be followed accordingly. Reliability of assay results
cannot be guaranteed if there are any deviations from the instructions in
In Vitro Diagnostic Medical Device this package insert.
NAME
Use By
Abbott RealTime HCV

Negative Control
INTENDED USE
The Abbott RealTime HCV assay is an in vitro reverse transcription-
polymerase chain reaction (RT‑PCR) assay for the quantitation of
Low Positive Control
hepatitis C viral ribonucleic acid (HCV RNA) in human serum and
plasma from HCV-infected individuals. The Abbott RealTime HCV assay
High Positive Control is intended for use as an aid in the management of HCV-infected
patients undergoing antiviral therapy. The Abbott RealTime HCV assay is
Calibrator A not for screening blood, plasma, serum or tissue donors for HCV, or to
be used as a diagnostic test to confirm the presence of HCV infection.
Calibrator B INTENDED USER
The intended users for the Abbott RealTime HCV assay are laboratory
and healthcare professionals.
Internal Control SUMMARY AND EXPLANATION OF THE TEST
HCV is a single-stranded RNA virus, with a genome of 9,500
nucleotides.1 HCV has been identified as the major etiological agent
Amplification Reagent Pack for post-transfusion non-A, non-B hepatitis worldwide.2,3 HCV has been
transmitted primarily through intravenous drug use and through blood
Upper Limit of Temperature products. Sensitive serological tests for HCV antibodies have greatly
reduced the incidence of new infections from donated blood. About 85%
of HCV-infected individuals develop chronic hepatitis, with up to 20%
of chronically infected individuals developing cirrhosis. In patients with
Consult instructions for use cirrhosis, the incidence of hepatocellular carcinoma is 1-4% per year.4,5
Quantitation of HCV RNA has been instrumental in understanding the
effectiveness of antiviral response to interferon monotherapy, interferon
Caution plus ribavirin combination therapy, and peginterferon plus ribavirin
combination therapy.6-10 Current guidelines for the management and
treatment of HCV recommend quantitative testing for HCV RNA before
the start of antiviral therapy, during therapy, and after the conclusion of
Warning treatment. The objective of treatment is a sustained virological response
(SVR), defined as the absence of HCV RNA detectable by a sensitive
test after the end of treatment. SVR is almost always preceded by an
Manufacturer early virological response (EVR), defined as a 2-log or greater decrease
in HCV viral load after 12 weeks of therapy.4,5,11
HCV RNA in serum or plasma can be quantitated using nucleic acid
Authorized Representative in the amplification or signal amplification technologies.12-14 The Abbott
European Community RealTime HCV assay uses RT-PCR technology combined with
See REAGENTS section for a full explanation of symbols used in homogeneous real time fluorescent detection for the quantitation of HCV
reagent component naming. RNA. The selection of a conserved region of the HCV genome provides
for the detection of genotypes 1 through 6. The assay is standardized
NOTICE TO USER against the Second WHO International Standard for Hepatitis C Virus
If a serious incident occurs in relation to this device, the incident should RNA (NIBSC Code 96/798)15 and results are reported in international
be reported to the manufacturer and to the appropriate competent units/mL (IU/mL).
authority of the member state in which the user and/or the patient is
established. To report to the manufacturer, see the contact information
BIOLOGICAL PRINCIPLES OF THE PROCEDURE
provided in the Customer service section or Technical assistance The Abbott RealTime HCV assay consists of 3 reagent kits:
section of these instructions. • Abbott RealTime HCV Amplification Reagent Kit
• Abbott RealTime HCV Control Kit
• Abbott RealTime HCV Calibrator Kit
The Abbott RealTime HCV assay uses RT-PCR16 to generate amplified
product from the RNA genome of HCV in clinical specimens. An RNA
sequence that is unrelated to the HCV target sequence is introduced
into each specimen at the beginning of sample preparation. This
unrelated RNA sequence is simultaneously amplified by RT-PCR and
1
serves as an internal control (IC) to demonstrate that the process hybridization to complementary sequences separates the fluorophore
has proceeded correctly for each sample. The amount of HCV target and the quencher and allows fluorescent emission and detection. The
sequence that is present at each amplification cycle is measured HCV and IC probes are each labeled with a different fluorophore, thus
through the use of fluorescent‑labeled oligonucleotide probes on the allowing for simultaneous detection of both amplified products at each
Abbott m2000rt instrument. The probes do not generate signal unless cycle. The amplification cycle at which fluorescent signal is detected
they are specifically bound to the amplified product. The amplification by the Abbott m2000rt is proportional to the log of the HCV RNA
cycle at which fluorescent signal is detected by the Abbott m2000rt is concentration present in the original sample.
proportional to the log of the HCV RNA concentration present in the PREVENTION OF NUCLEIC ACID CONTAMINATION
original sample.
The possibility of nucleic acid contamination is minimized because:
Sample Preparation • Reverse transcription, PCR amplification, and oligonucleotide
The purpose of sample preparation is to extract and concentrate the hybridization occur in a sealed Abbott 96-Well Optical
target RNA molecules, to make the target accessible for amplification, Reaction Plate.
and to remove potential inhibitors of amplification from the extract. • Detection is carried out automatically without the need to open the
The Abbott mSample Preparation System (4 × 24 Preps) uses magnetic Abbott 96-Well Optical Reaction Plate.
particle technology to capture nucleic acids and washes the particles • Pipettes with aerosol barrier tips or disposable transfer pipettes are
to remove unbound sample components. The bound nucleic acids are used for all pipetting. The disposable pipettes or pipette tips are
eluted and transferred to output tubes or a 96 deep-well plate. The discarded after use.
nucleic acids are then ready for amplification. The IC is taken through • Separate dedicated areas are used to perform the Abbott RealTime
the entire sample preparation procedure along with the calibrators, HCV assay. Refer to the SPECIAL PRECAUTIONS section of this
controls, and specimens. package insert.
Two automated instrument systems, the Abbott m2000sp or the Abbott
m1000 System can be used to prepare samples for the Abbott RealTime REAGENTS
HCV assay. The Abbott m2000sp provides automated sample eluate Abbott RealTime HCV Amplification Reagent Kit (List No.
transfer and reaction assembly in the Abbott 96-Well Optical Reaction 4J86-90)
Plate, whereas the Abbott m1000 System requires manual sample eluate 1. Abbott RealTime HCV Internal Control (List
transfer and reaction assembly. Alternatively, samples can be prepared No. 4J86Y) (4 vials, 1.2 mL per vial)
manually using the Abbott mSample Preparation System followed by < 0.01% noninfectious armored RNA with internal control sequences
manual reaction assembly. in negative human plasma. Negative human plasma tested and
Reagent Preparation and Reaction Plate Assembly found to be nonreactive for HBsAg, HBV DNA, HCV RNA, HIV RNA,
The Abbott m2000sp combines the Abbott RealTime HCV amplification anti‑HIV-1/HIV-2, and anti-HCV. Preservatives: 0.1% ProClin® 300 and
reagent components (HCV Oligonucleotide Reagent, Thermostable rTth 0.15% ProClin 950.
Polymerase Enzyme, and Activation Reagent). The Abbott m2000sp 2. Abbott RealTime HCV
dispenses the resulting master mix to the Abbott 96-Well Optical Amplification Reagent Pack (List No. 4J86) (4 packs, 24 tests/pack)
Reaction Plate along with aliquots of the nucleic acid samples prepared • 1 bottle (0.141 mL). Thermostable rTth Polymerase Enzyme (2.9
by the Abbott m2000sp. The plate is ready, after manual application of to 3.5 units/µL) in buffered solution.
the optical seal, for transfer to the Abbott m2000rt.
• 1 bottle (1.10 mL). HCV Oligonucleotide Reagent. < 0.1% synthetic
Abbott m1000 System users and manual sample preparation method oligonucleotides (4 primers and 2 probes) and < 0.3% dNTPs
users manually combine the Abbott RealTime HCV amplification reagent in a buffered solution with a reference dye. Preservatives: 0.1%
components to create the amplification master mix and transfer aliquots ProClin 300 and 0.15% ProClin 950.
of the master mix and sample eluates to the reaction plate. The plate is
ready, after manual application of the optical seal and centrifugation, for • 1 bottle (0.40 mL). Activation Reagent. 30 mM manganese
transfer to the Abbott m2000rt. chloride solution. Preservatives: 0.1% ProClin 300 and 0.15%
ProClin 950.
Amplification
Abbott RealTime HCV Control Kit (List No. 4J86-80)
During the amplification reaction on the Abbott m2000rt, the target
RNA is converted to cDNA by the reverse transcriptase activity of the 1. Abbott RealTime HCV Negative Control
thermostable rTth DNA polymerase. First, the HCV and IC reverse (List No. 4J86Z)(8 vials, 1.8 mL per vial)
primers anneal to their respective targets and are extended during a Negative human plasma tested and found to be nonreactive for
prolonged incubation period. After a denaturation step, in which the HBsAg, HBV DNA, HCV RNA, HIV RNA, anti-HIV-1/HIV-2, and anti-
temperature of the reaction is raised above the melting point of the HCV. Preservatives: 0.1% ProClin 300 and 0.15% ProClin 950.
double-stranded cDNA:RNA product, a second primer anneals to the 2. Abbott RealTime HCV Low Positive Control
cDNA strand and is extended by the DNA polymerase activity of the rTth (List No. 4J86W) (8 vials, 1.3 mL per vial)
enzyme to create a double-stranded DNA product. Noninfectious armored RNA with HCV sequences in negative human
During each round of thermal cycling, amplification products dissociate plasma. Negative human plasma tested and found to be nonreactive
to single strands at high temperature, allowing primer annealing and for HBsAg, HBV DNA, HCV RNA, HIV RNA, anti-HIV-1/HIV-2, and anti-
extension as the temperature is lowered. Exponential amplification of HCV. Preservatives: 0.1% ProClin 300 and 0.15% ProClin 950.
the product is achieved through repeated cycling between high and low 3. Abbott RealTime HCV High Positive Control
temperatures, resulting in a billion-fold or greater amplification of target (List No. 4J86X) (8 vials, 1.3 mL per vial)
sequences. Amplification of both targets (HCV and IC) takes place Noninfectious armored RNA with HCV sequences in negative human
simultaneously in the same reaction. plasma. Negative human plasma tested and found to be nonreactive
The target sequence for the Abbott RealTime HCV assay is in the 5´UTR for HBsAg, HBV DNA, HCV RNA, HIV RNA, anti-HIV-1/HIV-2, and
region of the HCV genome. This region is specific for HCV and is highly anti‑HCV. Preservatives: 0.1% ProClin 300 and 0.15% ProClin 950.
conserved.17 The primers are designed to hybridize to the 5´UTR region
Abbott RealTime HCV Calibrator Kit (List No. 4J86-70)
with the fewest possible mismatches among HCV genotypes 1 through 6.
1. Abbott RealTime HCV Calibrator A
The IC target sequence is derived from the hydroxypyruvate reductase
(List No. 4J86A) (12 vials, 1.3 mL per vial)
gene from the pumpkin plant, Cucurbita pepo, and is delivered in an
Noninfectious armored RNA with HCV sequences in negative human
Armored RNA® particle that has been diluted in negative human plasma.
plasma. Negative human plasma tested and found to be nonreactive
Detection for HBsAg, HBV DNA, HCV RNA, HIV RNA, anti-HIV-1/HIV-2, and anti-
During the read cycles of amplification on the Abbott m2000rt, the HCV. Preservatives: 0.1% ProClin 300 and 0.15% ProClin 950.
temperature is lowered further to allow fluorescent detection of 2. Abbott RealTime HCV Calibrator B
amplification products as the HCV and IC probes anneal to their targets (List No. 4J86B)(12 vials, 1.3 mL per vial)
(real-time fluorescence detection). The HCV and IC probes are single- Noninfectious armored RNA with HCV sequences in negative human
stranded DNA oligonucleotides consisting of a probe sequence with a plasma. Negative human plasma tested and found to be nonreactive
fluorescent moiety that is covalently linked to the 5´ end of the probe and for HBsAg, HBV DNA, HCV RNA, HIV RNA, anti-HIV-1/HIV-2, and
a quenching moiety that is covalently linked to the 3´ end of the probe. anti-HCV. Preservatives: 0.1% ProClin 300 and
In the absence of the HCV or IC target sequences, probe fluorescence 0.15% ProClin 950.
is quenched. In the presence of HCV or IC target sequences, probe
2
WARNINGS AND PRECAUTIONS aseptic technique should always be used when working with RNA.
Amplification technologies such as PCR are sensitive to accidental
introduction of product from previous amplification reactions. Incorrect
For In Vitro Diagnostic Use results could occur if either the clinical specimen or the RealTime
The Abbott RealTime HCV assay is not for screening blood, plasma, reagents used in the amplification step become contaminated by
serum or tissue donors for HCV, or to be used as a diagnostic test to accidental introduction of even a few molecules of amplification product.
confirm the presence of HCV infection. Measures to reduce the risk of contamination in the laboratory include
Safety Precautions physically separating the activities involved in performing PCR in
Refer to the Abbott m1000 Operating Manual, Safety Section, the Abbott compliance with good laboratory practices.
m2000sp and Abbott m2000rt Operations Manuals, Hazard Section, Work Areas
or the Manual Sample Preparation for Abbott RealTime RNA Assays, Use 3 dedicated areas within the laboratory for performing the Abbott
Handling Precaution section, for instructions on safety precautions. RealTime HCV assay with the Abbott m1000 System, or manual
sample preparation using the Abbott mSample Preparation System
CAUTION: This preparation contains human sourced and/or and the Abbott m2000rt:
potentially infectious components. Components sourced from human • The Reagent Preparation Area is dedicated to combining the Abbott
blood have been tested and found to be nonreactive by FDA-licensed RealTime HCV amplification reagent components to create the
tests for antibody to HCV, antibody to HIV-1, antibody to HIV-2, and amplification master mix and transferring aliquots of the master mix
HBsAg. The material is also tested and found to be negative by FDA- to the reaction plate. Laboratory coats, pipettes, pipette tips, and
licensed PCR methods for HIV-1 RNA and HCV RNA. No known test vortexers used in the Reagent Preparation Area must remain in this
method can offer complete assurance that products derived from human area and not be moved to either the Sample Preparation Area or
sources or inactivated microorganisms will not transmit infection. These the Amplification Area.
reagents and human specimens should be handled as if infectious • The Sample Preparation Area is dedicated to processing samples
using safe laboratory procedures, such as those outlined in Biosafety (specimens, Abbott RealTime HCV Controls, and Calibrators), and
in Microbiological and Biomedical Laboratories,18 OSHA Standards to adding processed samples, controls, and calibrators to the Abbott
on Bloodborne Pathogens,19 CLSI Document M29-A3,20 and other 96-Well Optical Reaction Plate. All reagents used in the Sample
appropriate biosafety practices.21 Therefore all human sourced materials Preparation Area should remain in this dedicated area at all times.
should be considered infectious. Laboratory coats, pipettes, pipette tips, and vortexers used in
These precautions include, but are not limited to, the following: the Sample Preparation Area must remain in this area and not be
• Wear gloves when handling specimens or reagents. moved to either the Reagent Preparation Area or the Amplification
• Do not pipette by mouth. Area. Do not bring amplification product into the Sample
Preparation Area.
• Do not eat, drink, smoke, apply cosmetics, or handle contact lenses
in areas where these materials are handled. • The Amplification Area is dedicated to the amplification and
detection of amplified product. Laboratory coats and equipment
• Clean and disinfect spills of specimens by including the use of a
used in the Amplification Area must remain in this area and not
tuberculocidal disinfectant such as 1.0% sodium hypochlorite or
be moved to either the Reagent Preparation Area or the Sample
other suitable disinfectant.18
Preparation Area.
• Decontaminate and dispose of all potentially infectious materials in
Only 2 dedicated areas, Sample Preparation Area and Amplification
accordance with local, state, and federal regulations.21
Area, are recommended when the Abbott m2000sp and Abbott
Components of the Abbott RealTime HCV Calibrator Kit (List No. 4J86- m2000rt are used.
70), the Abbott RealTime HCV Control Kit (List No. 4J86-80), and the
Components contained within a kit are intended to be used together.
Abbott RealTime HCV Amplification Reagent Kit (List No. 4J86-90)
Do not mix components from different kit lots. For example, do not use
contain the following components:
the negative control from control kit lot X with the positive controls from
• 2-Methyl-2H-isothiazol-3-one control kit lot Y.
• Reaction mass of: 5-chloro-2-methyl-4-isothiazolin-3-one Do not use kits or reagents after the dates shown on kit labels.
(EC no. 247-500-7) and 2-methyl-2H-isothiazol-3-one
Work areas and instrument platforms must be considered potential
(EC no. 220-239-6)(3:1)
sources of contamination. Change gloves after contact with potential
• Reaction mass of: 5-chloro-2-methyl-4-isothiazolin-3-one contaminants (specimens, eluates, and/or amplified product) before
(EC no. 247-500-7) and 2-methyl-4-isothiazolin-3-one handling unopened reagents, negative control, positive controls,
(EC no. 220-239-6)(3:1) calibrators, or specimens. Refer to the Abbott m1000 Operating Manual
The following warnings apply: and the Abbott m2000sp and Abbott m2000rt Operations Manuals for
Warning instrument cleaning procedures.
H317 May cause an allergic skin reaction. If the Abbott m1000 System or Abbott m2000sp instrument run is
aborted, dispose of all commodities and reagents according to the
P261 Avoid breathing mist/vapours/spray. Abbott m1000 Operating Manual or Abbott m2000sp Operations Manual.
P272 Contaminated work clothing should not be If the Abbott m2000sp master mix addition protocol is aborted, seal
allowed out of the workplace. the Abbott 96-Well Optical Reaction Plate in a sealable plastic bag and
P280 Wear protective gloves/protective clothing/ dispose according to the Abbott m2000sp Operations Manual, Hazards
eye protection. section, along with the gloves used to handle the plate.
P302+P352 IF ON SKIN: Wash with plenty of water. If the Abbott m2000rt instrument run is interrupted or aborted, seal the
P333+P313 If skin irritation or rash occurs: Get medical Abbott 96-Well Optical Reaction Plate in a sealable plastic bag and
advice/attention. dispose according to the Abbott m2000rt Operations Manual along with
the gloves used to handle the plate.
P362+P364 Take off contaminated clothing and wash
Decontaminate and dispose of all potentially biohazardous materials
before reuse.
in accordance with local, state, and federal regulations.21 All materials
P501 Dispose of contents / container in should be handled in a manner that minimizes the chance of potential
accordance with local regulations. contamination of the work area.
SPECIAL PRECAUTIONS NOTE: Autoclaving the sealed Reaction Plate will not degrade the
amplified product and may contribute to the release of the
Handling Precautions amplified product by opening the sealed plate. The laboratory
The Abbott RealTime HCV assay is only for use with human serum area can become contaminated with amplified product if the
and plasma specimens that have been handled and stored in capped waste materials are not carefully handled and contained before
tubes as described in the SPECIMEN COLLECTION, STORAGE, AND and after processing.
TRANSPORT TO THE TEST SITE section.
Aerosol Containment
During preparation of samples, compliance with good laboratory
practices is essential to minimize the risk of cross-contamination To reduce the risk of nucleic acid contamination due to aerosols formed
between samples, and the inadvertent introduction of ribonucleases during manual pipetting, aerosol barrier pipette tips must be used for
(RNases) into samples during and after the extraction procedure. Proper
3
all manual pipetting. The pipette tips must be used only 1 time. Clean Freshly drawn specimens (whole blood) may be held at 2 to 30°C for up
and disinfect spills of specimens and reagents as stated in the Abbott to 6 hours prior to centrifugation.
m1000 Operating Manual or the Abbott m2000sp and Abbott m2000rt Separate serum or plasma from cells by centrifugation.
Operations Manuals. After centrifugation, serum or plasma may be removed from cells. Serum
Contamination and Inhibition or plasma specimens may be stored:
The following precautions should be observed to minimize the risks • at 15 to 30°C for up to 24 hours
of RNase contamination, cross‑contamination between samples, and • at 2 to 8°C for up to 3 days
inhibition: • at – 70°C or colder for longer term22-25
• Wear appropriate personal protective equipment at all times. Multiple freeze/thaw cycles should be avoided. If frozen, thaw
• Use powder-free gloves. specimens at 15 to 30°C or at 2 to 8°C. Once thawed, if specimens are
• Change gloves after having contact with potential contaminants not being processed immediately, they can be stored at 2 to 8°C for up
(specimens, eluates, and/or amplified product). to 6 hours.
• To reduce the risk of nucleic acid contamination due to aerosols NOTE: Serum and plasma specimens should not be frozen in non-gel
formed during pipetting, pipettes with aerosol barrier tips must be blood collection tubes.
used for all pipetting. The length of the tip should be sufficient to Specimen Transport
prevent contamination of the pipette barrel. While pipetting, care
Ship specimens according to recommended storage temperature and
should be taken to avoid touching the pipette barrel to the inside of
time listed in the Specimen Collection and Storage section above.
the sample tube or container. The use of extended aerosol barrier
For domestic and international shipments, specimens should be
pipette tips is recommended.
packaged and labeled in compliance with applicable state, federal, and
• Change aerosol barrier pipette tips between ALL manual liquid international regulations
transfers.
covering the transport of clinical, diagnostic, or biological specimens.
• The Abbott m Sample Preparation System (4 × 24 Preps) reagents
are single use only. Use new reagent troughs or vessels, reaction ABBOTT REALTIME HCV ASSAY PROCEDURE
vessels, and newly opened reagents for every new Abbott RealTime This Abbott RealTime HCV package insert contains 2 assay protocols:
HCV assay run. At the end of each run, discard all remaining • Samples prepared for amplification using the Abbott m1000
reagents from the worktable as stated in the Abbott m1000 System, or the manual sample preparation method, follow ASSAY
Operating Manual or the Abbott m2000sp Operations Manual and PROTOCOL I.
the Abbott mSample Preparation System (4 × 24 Preps) product • Samples prepared for amplification using the Abbott m2000sp
information sheet. instrument follow ASSAY PROTOCOL II.
STORAGE INSTRUCTIONS The Abbott RealTime HCV assay provides 2 sample volume options
(0.2 mL and 0.5 mL).
Abbott RealTime HCV Amplification Reagent Kit
(List No. 4J86-90). (See assay protocol step 6 and INTERPRETATION OF RESULTS
section.)
The Abbott RealTime HCV Amplification Reagent Pack and
Internal Control vials must be stored at – 10°C or colder Materials Provided
when not in use. Care must be taken to separate the Abbott • Abbott RealTime HCV Amplification Reagent Kit (List No. 4J86-90)
RealTime HCV Amplification Reagent Pack that is in use from Materials Required But Not Provided
direct contact with samples, calibrators, and controls.
• Abbott RealTime HCV Calibrator Kit (List No. 4J86-70)
Abbott RealTime HCV Control Kit (List No. 4J86-80). • Abbott RealTime HCV Control Kit (List No. 4J86-80)
The Abbott RealTime HCV Negative and Positive Controls must For manual sample preparation, refer to the Materials and Equipment
be stored at – 10°C or colder. Required Section of the Manual Sample Preparation for Abbott RealTime
RNA Assays procedure (List No. 06L73).
Abbott RealTime HCV Calibrator Kit (List No. 4J86-70).
For Abbott m1000 System For Abbott m2000sp Instrument
The Abbott RealTime HCV Calibrator A and Calibrator B must
be stored at – 10°C or colder. Sample Preparation Area Sample Preparation Area
• Abbott m1000 System • Abbott m2000sp instrument
SHIPPING CONDITIONS • Reaction Vessels • 5 mL Reaction Vessels
• Abbott RealTime HCV Amplification Reagent Kit: Ship on dry ice. • Abbott mSample Preparation • Abbott mSample Preparation
• Abbott RealTime HCV Control Kit: Ship on dry ice. System (4 × 24 Preps) System (4 × 24 Preps)
(List No. 04J70-24) (List No. 04J70-24)
• Abbott RealTime HCV Calibrator Kit: Ship on dry ice.
• Calibrated precision pipettes • Calibrated precision pipettes
INDICATION OF INSTABILITY OR DETERIORATION capable of delivering 20 to capable of delivering 20 to
OF REAGENTS 1000 µL 1000 µL
When a positive or negative control value is out of the expected range, • Aerosol barrier pipette tips for • Aerosol barrier pipette tips for 20
it may indicate deterioration of the reagents. Associated test results 20 to 1000 µL pipettes to 1000 µL pipettes
are invalid and samples must be retested. Assay recalibration may • 11.6 to 16 mm sample tubes • 13 to 16 mm sample tubes
be necessary. • 200 µL and 1000 µL disposable • 200 µL and 1000 µL disposable
INSTRUMENT PROCEDURE tips tips
The nucleic acid testing (NAT) software must be installed on the Abbott • Abbott 96 Deep-Well Plate • Vortex mixer
m1000 System prior to performing the assay. For detailed information on (List No. 04J71-30)
NAT software installation, refer to the Abbott m1000 Operating Manual,
Putting into Operation section.
The Abbott RealTime HCV application files must be installed on the
Abbott m2000sp and Abbott m2000rt systems from the Abbott RealTime
HCV m2000 ROW System Combined Application CD-ROM prior to
performing the assay. For detailed information on application file
installation, refer to the Abbott m2000sp and Abbott m2000rt Operations
Manuals, Operating Instructions section.
SPECIMEN COLLECTION, STORAGE, AND TRANSPORT
TO THE TEST SITE
Specimen Collection and Storage
Human serum and plasma (EDTA and ACD-A) specimens may be
used with the Abbott RealTime HCV assay. Follow the manufacturer’s
instructions for processing collection tubes.

4
• Vortex mixer • Abbott Optical Adhesive Covers ASSAY PROTOCOL I: ABBOTT m1000 SYSTEM OR
• Abbott Optical Adhesive Covers (List No. 04J71-75) THE MANUAL SAMPLE PREPARATION METHOD, AND
(List No. 04J71-75) • Abbott Adhesive Cover Applicators ABBOTT m2000rt INSTRUMENT
• Abbott Adhesive Cover • Abbott Splash-Free Support Base For a detailed description of how to perform an Abbott m1000 System
Applicators (List No. 09K31-01) and Abbott m2000rt protocol refer to the Abbott m1000 Operating
• Abbott Splash-Free Support • Master Mix Vial Manual, Operation section, and the Abbott m2000rt Operations Manual,
Base (List No. 09K31-01) Operating Instructions section.
• 200 mL reagent vessels
• Reagent troughs Laboratory personnel must be trained to operate the Abbott m1000
• Abbott 96-Deep-Well Plate
System and the Abbott m2000rt instrument. The operator must have a
• 1.5 mL output tubes (List No. 04J71-30)
thorough knowledge of the software applications and must follow good
• Centrifuge capable of 5,000g • Abbott RealTime HCV m2000 laboratory practices.
ROW System Combined 1. Thaw assay controls and IC at 15 to 30°C or at 2 to 8°C. Thaw
Application CD‑ROM calibrators at 15 to 30°C or at 2 to 8°C only if performing a
(List No. 1L69) calibration run; see QUALITY CONTROL PROCEDURES section of
• Abbott 96-Well Optical Reaction this package insert.
Plate (List No. 04J71-70)
• Once thawed, assay controls, IC, and calibrators can be stored at
• Centrifuge capable of 2,000g 2 to 8°C for up to 24 hours before use.
For Abbott m1000 System For Abbott m2000rt Instrument • Vortex each assay calibrator and each control 3 times for 2 to 3
seconds before use. Ensure that the contents of each vial are at
Reagent Preparation Area Amplification Area
the bottom after vortexing by tapping the vials on the bench to
• PCR cooler, either • Abbott m2000rt instrument bring liquid to the bottom of the vial.
StrataCooler® 96 Benchtop • Abbott RealTime HCV m2000 2. Thaw amplification reagents at 15 to 30°C or at 2 to 8°C and store at
Cooler or Eppendorf ROW System Combined 2 to 8°C until required for the amplification master mix procedure.
PCR-Cooler Application CD-ROM
• Abbott 96-Well Optical (List No. 1L69) • Once thawed, the amplification reagents can be stored at 2 to
Reaction Plate 8°C for up to 24 hours if not used immediately.
• Abbott m2000rt Optical
(List No. 04J71-70) Calibration Kit NOTE: Use 1 bottle of mLysis Buffer, 1 vial of IC, and 1 Abbott
• Calibrated precision pipettes (List No. 04J71-93) RealTime HCV Amplification Reagent Pack to support up
capable of delivering 20 to to 24 reactions. Use a second set of reagents to support
1000 µL 25 to 48 reactions. A maximum of 48 reactions can be
performed per run using an Abbott m1000 instrument.
• Aerosol barrier pipette tips for
20 to 1000 µL pipettes Sample Preparation Area
• Single-use RNase/DNase-free For sample preparation using the Abbott m1000 System, follow steps
tube or container 3 through 10. For the manual sample preparation method refer to
• Vortex mixer Extraction Protocol Section of the Manual Sample Preparation for Abbott
RealTime RNA Assays procedure.
Other Materials
3. Gently invert the Abbott mSample Preparation bottles to ensure a
• Biological safety cabinet approved for working with infectious
homogeneous solution. If crystals are observed in any of the reagent
materials
bottles upon opening, allow the reagent to equilibrate at room
• Sealable plastic bags temperature until the crystals disappear. Do not use the reagents
• RNase-free water (Eppendorf or equivalent)* until the crystals have dissolved.
• 1.7 mL molecular biology grade microcentrifuge tubes (Dot 4. Vortex each IC 3 times for 2 to 3 seconds before use.
Scientific, Inc. or equivalent)*
5. Use a calibrated precision PIPETTE DEDICATED FOR INTERNAL
• Cotton tip applicators (Puritan or equivalent)*
*NOTE: These 3 items are used in the procedure for Monitoring the
CONTROL USE ONLY to add 500 µL of IC to each bottle of mLysis
Buffer. Mix by gently inverting the container 5 to 10 times to minimize
Laboratory for the Presence of Contamination. Refer to the foaming.
QUALITY CONTROL PROCEDURES section of this package
insert. 6. A total of 48 samples can be processed in each run. A negative
control, a low positive control, and a high positive control are
Procedural Precautions included in each run, therefore allowing a maximum of 45 specimens
Read the instructions in this package insert carefully before processing to be processed per run.
samples. • The Abbott RealTime HCV assay minimum sample volume and
The Abbott RealTime HCV Calibrators, Internal Control, Negative Control, associated rack requirements on the Abbott m1000 System are:
and Low and High Positive Control vials are intended for single-use only
and should be discarded after use. Abbott RealTime HCV
Use aerosol barrier pipette tips or disposable pipettes only 1 time Minimum Sample Volume
when pipetting specimens, IC, or amplification reagents. To prevent Assay Application
contamination to the pipette barrel while pipetting, care should be Rack Tube Diametera 0.2 mL 0.5 mL
taken to avoid touching the pipette barrel to the inside of the sample 13 mm 11.6 mm - 14.0 mm 0.7 mL 1.0 mL
tube or container. The use of extended aerosol barrier pipette tips is
recommended. 16 mm 15.0 mm - 16.0 mm 1.0 mL 1.3 mL
a Refers to sample tube outer diameter
Monitoring procedures for the presence of amplification product can be
found in the QUALITY CONTROL PROCEDURES section in this package • If frozen, thaw specimens at 15 to 30°C or at 2 to 8°C. Once
insert. thawed, specimens can be stored at 2 to 8°C for up to 6 hours if
To reduce the risk of nucleic acid contamination, clean and disinfect not processed immediately.
spills of specimens by including the use of a tuberculocidal disinfectant • Vortex each specimen 3 times for 2 to 3 seconds before
such as 1.0% sodium hypochlorite or other suitable disinfectant. loading on the Abbott m1000 System worktable. Specimens
The Abbott RealTime HCV Calibrators and Controls must be prepared showing particulate matter or turbidity should be clarified by
in conjunction with the specimens to be tested. The use of the Abbott centrifugation at 2,000g for 5 minutes prior to testing. Aliquot
RealTime HCV Controls and Calibrators is integral to the performance each specimen into clean tubes or vials if necessary. Refer to the
of the Abbott RealTime HCV assay. Refer to the QUALITY CONTROL Abbott m1000 Operating Manual for tube sizes. Avoid touching the
PROCEDURES section of this package insert for details. inside of the cap when opening tubes.
7. Place the calibrators (if applicable), the low and high positive
controls, the negative control, and the patient specimens into the
Abbott m1000 sample rack. Follow directions for performing a
user-defined protocol, as described in the Abbott m1000 Operating
Manual, Operation section.
5
8. Place the Reaction Vessels into the Abbott m1000 1 mL 19. Remove the Abbott 96-Well Optical Reaction Plate from the PCR
subsystem carrier. cooler to the Abbott Splash-Free Support Base. Centrifuge the
9. Load the Abbott mSample Preparation System reagents and the Abbott 96-Well Optical Reaction Plate in the Abbott Splash-Free
1.5 mL Output Tubes on the Abbott m1000 System worktable as Support Base at 5000g for 5 minutes. Transfer to the Amplification
described in the Abbott m1000 Operating Manual, Operation section. Area.
10. Initiate the Abbott m1000 protocol as described in the Abbott m1000 NOTE: Do not transfer the PCR cooler to the Amplification Area.
Operating Manual, Operation Section. From the Protocol screen, Amplification Area
select the appropriate application file corresponding to the sample 20. Place the Abbott 96-Well Optical Reaction Plate in the Abbott
volume being tested. m2000rt instrument. From the Protocol screen, select the appropriate
• The assembly of the amplification master mix and sample application file corresponding to the sample volume being tested.
eluates into the Abbott 96-Well Optical Reaction Plate (step 17) Initiate the Abbott RealTime HCV protocol, as described in the Abbott
must be initiated within 1 hour after completion of m2000rt Operations Manual, Operating Instructions Section.
Sample Preparation.
POST PROCESSING PROCEDURES
Amplification Area 1. Clean the PCR cooler as described in the PCR cooler instruction
11. Switch on and initialize the Abbott m2000rt instrument. manual and return to the Reagent Preparation Area.
NOTE: The Abbott m2000rt instrument requires 15 minutes to 2. Remove the 1.5 mL Output Tubes from the worktable and dispose of
warm up. according to the Abbott m1000 Operating Manual.
12. Create the Abbott m2000rt test order. Refer to the Operating 3. Place the Abbott 96-Well Optical Reaction Plate in a sealable plastic
Instructions section of the Abbott m2000rt Operations Manual. bag and dispose of according to the Abbott m2000rt Operations
From the Protocol screen, select the appropriate application file Manual along with the gloves used to handle the plate.
corresponding to the sample volume being tested.
4. Clean the Splash-Free Support Base before next use, according to
• Enter calibrator (needed if a calibration curve has not been stored the Abbott m2000rt Operations Manual.
on the Abbott m2000rt) and control lot specific values in the test
5. For the manual sample preparation method users, refer to the Clean
order for accurate calibration and control evaluation. Lot-specific
Up section of the Manual Sample Preparation for Abbott RealTime
values are specified in each Abbott RealTime HCV Calibrator and
RNA Assays Procedure (List No. 06L73).
Control Kit Card.
Reagent Preparation Area ASSAY PROTOCOL II: ABBOTT m2000sp INSTRUMENT
All reagent preparation must take place in the dedicated Reagent AND ABBOTT m2000rt INSTRUMENT
Preparation Area. Refer to the Handling Precautions section of this For a detailed description of how to perform an Abbott m2000sp
package insert before preparing reagents. instrument and Abbott m2000rt instrument protocol refer to the
NOTE: Change gloves before handling the amplification reagents. Abbott m2000sp and Abbott m2000rt Operations Manuals, Operating
13. Prepare the amplification master mix. Instructions sections. The 96-sample capability requires Abbott
m2000sp Software Version 4.0 or higher. Please follow Abbott m2000sp
• Each amplification reagent pack supports up to 24 reactions. Operations Manual (List 09K20-04 or higher) and addendum or
• Prior to opening the amplification reagents, ensure that the addenda.
contents of the vials are at the bottom by tapping the vials in an Laboratory personnel must be trained to operate the Abbott m2000sp
upright position on the bench to bring the liquid to the bottom of and Abbott m2000rt instruments. The operator must have a thorough
the vials. knowledge of the applications run on the instruments and must follow
• Prepare the master mix by using a PIPETTE DEDICATED FOR good laboratory practices.
REAGENT USE ONLY to add 271 µL of the HCV Activation 1. Thaw assay controls and IC at 15 to 30°C or at 2 to 8°C. Thaw
Reagent (Reagent 1) and 949 µL of the HCV Oligonucleotide calibrators at 15 to 30°C or at 2 to 8°C only if performing a
Reagent (Reagent 2) together in the Thermostable rTth DNA calibration run; see QUALITY CONTROL PROCEDURES section of
Polymerase Enzyme bottle (Reagent 3). this package insert.
• If performing 25 to 48 reactions, prepare a second amplification • Once thawed, assay controls, IC, and calibrators can be stored at
master mix with a second amplification reagent pack. 2 to 8°C for up to 24 hours before use.
• The Abbott m2000rt protocol (step 20) must be initiated within • Vortex each assay calibrator and each control 3 times for 2 to 3
40 minutes of the addition of Activation Reagent into the first seconds before use. Ensure that the contents of each vial are at
rTth Enzyme Reagent bottle (step 13). the bottom after vortexing by tapping the vials on the bench to
14. Pipette the contents of the master mix from the enzyme bottle(s) bring liquid to the bottom of the vial.
into a single-use RNase/DNase-free tube and vortex to mix. 2. Thaw amplification reagents at 15 to 30°C or at 2 to 8°C and store at
15. Place an Abbott 96-Well Optical Reaction Plate in a PCR cooler, 2 to 8°C until required for the amplification master mix procedure.
stored as indicated in the PCR cooler instruction manual. Using a • Once thawed the amplification reagents can be stored at 2 to 8°C
DEDICATED PIPETTE, dispense 50 µL aliquots of the amplification for up to 24 hours if not used immediately.
master mix into the Abbott 96-Well Optical Reaction Plate. A
NOTE: Use 1 bottle of mLysis Buffer, 1 vial of IC, and 1 Abbott
calibrated repeat pipettor may be used. Visually verify that 50 µL has
RealTime HCV Amplification Reagent Kit to support up to
been dispensed into each well.
24 reactions. Use a second set of reagents to support 25
16. Transfer the Abbott 96-Well Optical Reaction Plate on the PCR to 48 reactions, a third set of reagents to support 49 to 72
cooler to the Sample Preparation Area. reactions and a fourth set of reagents to support 73 to 96
Sample Preparation Area reactions WITH THE EXCEPTION OF mMICROPARTICLES.
USE ONLY TWO BOTTLES OF mMICROPARTICLES WHEN
17. In the Sample Preparation Area, transfer 50 µL of sample eluate to
PROCESSING 25 TO 96 REACTIONS.
the Abbott 96-Well Optical Reaction Plate on the PCR cooler. Use
a separate pipette tip for each sample eluate transfer. During the 3. Gently invert the Abbott mSample Preparation bottles to ensure a
transfer of each sample, mix the reaction by pipetting up and down homogeneous solution. If crystals are observed in any of the reagent
3 to 5 times. Visually verify that 100 µL has been dispensed into bottles upon opening, allow the reagent to equilibrate at room
each well. temperature until the crystals disappear. Do not use the reagents
until the crystals have dissolved.
18. Seal the Abbott 96-Well Optical Reaction Plate according to the
instructions in the Abbott m2000rt Operations Manual. 4. Vortex each IC 3 times for 2 to 3 seconds before use.
5. Use a calibrated precision PIPETTE DEDICATED FOR INTERNAL
CONTROL USE ONLY to add 500 µL of IC to each bottle of mLysis
Buffer. Mix by gently inverting the container 5 to 10 times to minimize
foaming.

6
6. A total of 96 samples can be processed in each run. A negative Buffer (Reagent Carrier 2, location 6). If this reagent vessel has
control, a low positive control, and a high positive control are been unloaded, place a new reagent vessel with the mElution
included in each run, therefore allowing a maximum of 93 specimens Buffer label into Reagent Carrier 2, location 6. System fluid will
to be processed per run. be added to the reagent vessel and used to fill empty wells. Once
• The Abbott RealTime HCV assay minimum sample volume and this process is complete, the system will continue with the master
associated rack requirements on the Abbott m2000sp are: mix addition.
NOTE: System instructions for use of the automated plate-filling
Abbott RealTime HCV feature are found in the Abbott m2000sp Operations
Minimum Sample Volume Manual (List No. 9K20-04 or higher), section 5, Operating
Assay Application Instructions, Sample Extraction—Closed Mode.
Rack Tube Diametera 0.2 mL 0.5 mL • The Abbott m2000rt protocol (step 16) must be started within
13 mm 11.6 mm - 14.0 mm 0.7 mL 1.0 mL 50 minutes of the initiation of the Master Mix Addition protocol
16 mm 15.0 mm - 16.0 mm 1.0 mL 1.3 mL (step 12).
a Refers to sample tube outer diameter NOTE: If the run is aborted for any reason subsequent to step 12,
a new 96-well PCR plate must be used if the Abbott
• If frozen, thaw specimens at 15 to 30°C or at 2 to 8°C. Once m2000sp Master Mix Addition Protocol (Step 12) will be
thawed, specimens can be stored at 2 to 8°C for up to 6 hours, if repeated.
not processed immediately. 13. Switch on and initialize the Abbott m2000rt instrument in the
• Vortex each specimen 3 times for 2 to 3 seconds before amplification area.
loading on the Abbott m2000sp worktable. Specimens
NOTE: The Abbott m2000rt requires 15 minutes to warm-up.
showing particulate matter or turbidity should be clarified by
centrifugation at 2,000g for 5 minutes prior to testing. Aliquot NOTE: Remove gloves before returning to the sample preparation
each specimen into clean tubes or vials if necessary. Refer to the area.
Abbott m2000sp Operations Manual for tube sizes. Avoid touching 14. Seal the Abbott 96-Well Optical Reaction Plate after the Abbott
the inside of the cap when opening tubes. m2000sp instrument has completed addition of samples and master
7. Place the low and high positive controls, the negative control, the mix according to the Abbott m2000sp Operations Manual, Operating
calibrators, if applicable, and the patient specimens into the Abbott Instructions section.
m2000sp sample rack. 15. Place the sealed optical reaction plate into the Splash-Free Support
8. Place the 5 mL Reaction Vessels into the Abbott m2000sp 1 mL Base for transfer to the Abbott m2000rt instrument.
subsystem carrier. 16. Place the Abbott 96-Well Optical Reaction Plate in the Abbott
9. Load the Abbott mSample Preparation System reagents and the m2000rt instrument. From the Protocol screen, select the appropriate
Abbott 96 Deep-Well Plate on the Abbott m2000sp worktable as application file corresponding to the sample volume being tested.
described in the Abbott m2000sp Operations Manual, Operating Initiate the Abbott RealTime HCV protocol, as described in the Abbott
Instructions. m2000rt Operations Manual, Operating Instructions section.
10. From the Protocol screen, select the appropriate application file NOTE: If creating the Abbott m2000rt test order manually, enter
corresponding to the sample volume being tested. Initiate the sample sample IDs in the corresponding PCR tray locations
extraction protocol as described in the Abbott m2000sp Operations according to the “Wells for Selected Plate” grid, found on
Manual, Operating Instruction. the detail screen of the “PCR Plate Results” on the Abbott
m2000sp. See Section 5 of the Abbott m2000sp
• Enter calibrator (needed if a calibration curve has not been stored Operations Manual.
on the Abbott m2000rt) and control lot specific values in the
Sample Extraction: Worktable Setup, Calibrator and Control POST PROCESSING PROCEDURES
fields. Lot-specific values are specified in each Abbott RealTime 1. Remove the Abbott 96 Deep-Well Plate from the worktable and
HCV Calibrator and Control Kit Card. dispose of according to the Abbott m2000sp Operations Manual.
• The Abbott m2000sp Master Mix Addition protocol (step 12) 2. Place the Abbott 96-Well Optical Reaction Plate in a sealable
must be initiated within 1 hour after completion of Sample plastic bag and dispose according to the Abbott m2000rt Operations
Preparation. Manual, along with the gloves used to handle the plate.
NOTE: Change gloves before handling the amplification reagents. 3. Clean the Splash-Free Support Base before next use, according to
11. Load the amplification reagents and the master mix vial on the the Abbott m2000rt Operations Manual.
Abbott m2000sp worktable after sample preparation is completed.
QUALITY CONTROL PROCEDURES
• Each amplification reagent pack supports up to 24 reactions.
• Prior to opening the amplification reagents, ensure that the Abbott m2000rt Optical Calibration
contents are at the bottom of the vials by tapping the vials in an Refer to the Calibration Procedures section in the Abbott m2000rt
upright position on the bench. Operations Manual for a detailed description of how to perform an
• Remove and discard the amplification vial caps. Abbott m2000rt Optical Calibration.
• A second amplification reagent pack is required if performing 25 Optical calibration of the Abbott m2000rt instrument is required for the
to 48 reactions. accurate measurement and discrimination of dye fluorescence during
the Abbott RealTime HCV assay.
• A third amplification reagent pack is required if performing 49 to
72 reactions. The following Abbott m2000rt Optical Calibration Plates are used to
calibrate the Abbott m2000rt instrument for the Abbott RealTime HCV
• A fourth amplification reagent pack is required if performing 73 to assay:
96 reactions.
• FAM™ Plate (Carboxyfluorescein)
12. Select the appropriate deep well plate that matches the
corresponding sample preparation extraction. Initiate the Abbott • ROX™ Plate (Carboxy-X-rhodamine)
m2000sp Master Mix Addition protocol. Follow the instructions as • VIC® Plate (Proprietary dye)
described in the Abbott m2000sp Operations Manual, Operating Assay Calibration
Instructions section. For a detailed description of how to perform an assay calibration refer to
NOTE: The operator should not manually fill any empty/unfilled the Abbott m2000sp and Abbott m2000rt Operations Manuals, Operating
wells in the Abbott 96-Well Optical Reaction Plate. Instructions sections.
• After sample extraction is complete, the Abbott m2000sp A calibration curve is required to quantitate the HCV RNA concentration
automatically fills any empty wells in the Abbott 96-Well Optical of specimens and controls. Two assay calibrators are run in replicates
Reaction Plate when there are greater than 48 samples of 3 to generate a calibration curve (HCV concentration versus the
processed within a run. Plate fill is not performed for runs threshold cycle [Ct] at which a reactive level of fluorescent signal is
containing 48 samples or fewer. detected). The calibration curve slope and intercept are calculated and
• If prompted by the instrument, Reagent Carrier 2 should remain stored on the instrument. The concentration of HCV RNA in a sample is
in place, minimally containing the reagent vessel for mElution calculated from the stored calibration curve. Results are automatically
reported on the Abbott m2000rt workstation.
7
Follow the procedure for sample extraction, master mix addition, 7. Cap the microcentrifuge tube.
amplification and detection protocols as stated in the Abbott m1000 8. Test this sample according to the assay procedure section of this
Operating Manual or Abbott m2000sp Operations Manual, and the package insert.
Abbott m2000rt Operations Manual.
• Transfer liquid from the microcentrifuge tube to a 5 mL Reaction
Once an Abbott RealTime HCV calibration is accepted and stored, it may
Vessel.
be used for 6 months. During this time, all subsequent samples may be
tested without further calibration unless: • Bring the volume to 1.5 mL with RNase-free water.
• An Abbott RealTime HCV Amplification Reagent Kit with a new lot 9. The presence of contamination is indicated by the detection of HCV
number is used. nucleic acid in the swab samples.
• An Abbott mSample Preparation System (4 × 24 Preps) with a new 10. If HCV nucleic acid is detected on equipment, follow the cleaning
lot number is used. and decontaminating guidelines given in that equipment’s operations
• An Abbott RealTime HCV application file for a different sample manual. If HCV nucleic acid is detected on surfaces, clean the
volume is used. contaminated areas with 1.0% (v/v) sodium hypochlorite solution,
followed by 70% ethanol or water.
Detection of Inhibition
NOTE: Chlorine solutions may pit equipment and metal. Use
An IC threshold cycle (Ct) assay validity parameter is established during sufficient amounts or repeated applications of 70% ethanol
a calibration run. or water until chlorine residue is no longer visible.
A defined, consistent quantity of IC is introduced into each specimen, 11. Repeat testing of the contaminated area by following steps 1
calibrator, and control at the beginning of sample preparation and through 10.
measured on the Abbott m2000rt instrument to demonstrate proper
specimen processing and assay validity. The IC is composed of an RNA RESULTS
sequence unrelated to the HCV target sequence. Calculation
The median amplification cycle at which the IC target sequence The concentration of viral HCV RNA in a sample or control is calculated
fluorescent signal is detected in calibration samples establishes an IC Ct from the stored calibration curve. The Abbott m2000rt instrument
validity range to be met by all subsequent processed specimens. automatically reports the results on the Abbott m2000rt workstation.
An error control flag is displayed when a specimen or control fails to Assay results can be reported in IU/mL or Log IU/mL.
meet this specification. Refer to the Abbott m2000rt Operations Manual
for an explanation of the corrective actions for the error control flag. INTERPRETATION OF RESULTS
Specimens whose IC Ct value exceeds the established range must be Sample Volume Result Interpretation
retested starting with sample preparation. 0.5 mL Not Detected Target not detected
Negative and Positive Controls < 1.08 Log IU/mLa Detected
A negative control, a low positive control, and a high positive control are 1.08 to 8.00 Log IU/mL
included in each test order to evaluate run validity.
> 8.00 Log IU/mL > ULQc
The lot specific values for the low positive control and high positive
control are specified on each Abbott RealTime HCV Control Kit Card and
must be entered into the assay test order when a run is performed. 0.2 mL Not Detected Target not detected
An error control flag is displayed when a control result is out of range. < 1.48 Log IU/mLb Detected
Refer to the Abbott m2000rt Operations Manual for an explanation of 1.48 to 8.00 Log IU/mL
the corrective actions for the error control flag. If negative or positive > 8.00 Log IU/mL > ULQ
controls are out of range, all of the specimens and controls from that
run must be reprocessed, beginning with sample preparation. a 12 IU/mL
b 30 IU/mL
The presence of HCV must not be detected in the negative control. HCV
detected in the negative control is indicative of contamination by other c ULQ = upper limit of quantitation
samples or by amplified product introduced during sample preparation LIMITATIONS OF THE PROCEDURE
or during preparation of the Abbott 96-Well Optical Reaction Plate.
• FOR IN VITRO DIAGNOSTIC USE
To avoid contamination, clean the Abbott m1000 System or Abbott
m2000sp instrument and the Abbott m2000rt instrument and repeat • Optimal performance of this test requires appropriate specimen
sample processing for controls and specimens following the Procedural collection, handling, preparation, and storage (refer to the
Precautions. If negative controls are persistently reactive, contact your SPECIMEN COLLECTION, STORAGE, AND TRANSPORT TO THE
Abbott representative. TEST SITE section of this package insert).
• Human serum and plasma specimens (ACD-A or EDTA) may
Monitoring the Laboratory for the Presence of Contamination be used with the Abbott RealTime HCV assay. The use of other
It is recommended that this test be done at least once a month to anticoagulants has not been validated with the Abbott RealTime HCV
monitor laboratory surfaces and equipment for contamination by assay.
amplification product. It is very important to test all areas that may have • Use of the Abbott RealTime HCV assay is limited to personnel who
been exposed to processed specimens, controls, and calibrators, and/ have been trained in the procedures of a molecular diagnostic assay
or amplification product. This includes routinely handled objects such and the Abbott m1000 System, Abbott m2000sp, and Abbott
as pipettes, the Abbott m1000 System, the Abbott m2000sp and Abbott m2000rt instruments.
m2000rt function keys, laboratory bench surfaces, microcentrifuges, and
• The instruments and assay procedures reduce the risk of
centrifuge adaptors.
contamination by amplification product. However, nucleic acid
1. Add 0.8 mL RNase-free water to a 1.7 mL molecular biology grade contamination from the calibrators, positive controls, or specimens
microcentrifuge tube. must be controlled by good laboratory practice and careful
2. Saturate the cotton tip of an applicator (Puritan or equivalent) in the adherence to the procedures specified in this package insert.
RNase-free water from the microcentrifuge tube. • As with any diagnostic test, results from the Abbott RealTime HCV
3. Using the saturated cotton tip of the applicator, wipe the area to be assay should be interpreted in conjunction with other clinical and
monitored using a sweeping motion. Place the applicator into the laboratory findings. A specimen with the result of “Not Detected”
microcentrifuge tube. cannot be presumed to be negative for HCV RNA.
4. Swirl the cotton tip in RNase-free water 10 times, and then press the SPECIFIC PERFORMANCE CHARACTERISTICS
applicator along the inside of the tube so that the liquid drains back The following performance characteristics were determined using the
into the solution at the bottom of the microcentrifuge tube. Discard Abbott RealTime HCV assay with the Abbott m2000sp and the 0.5 mL
the applicator. sample preparation procedure, unless otherwise specified.
5. Pipette 0.5 mL of mWash 1 buffer to a clean tube using the pipette Limit of Detection (LOD)
dedicated for Internal Control use. The LOD is defined as the HCV RNA concentration detected with a
6. Add 20 µL of the mWash 1 buffer to each microcentrifuge tube. probability of 95% or greater.

8
Limit of Detection, 0.5 mL Sample Volume Figure 1. Plasma
The LOD of the Abbott RealTime HCV assay is 12 IU/mL with the 0.5 mL
sample preparation procedure.
The LOD was determined by testing dilutions of the Second WHO
International Standard for Hepatitis C Virus RNA (NIBSC 96/798)
prepared in HCV negative human plasma. Testing was performed with
3 lots of amplification reagents on 3 instrument systems. The results,
representative of the analytical sensitivity of the Abbott RealTime HCV
assay, are summarized in Table 1.
Table 1.
Number Number Percent
(IU/mL) Tested Detected Detected
25.0 57 57 100
20.0 57 57 100
15.0 57 55 96
12.5 57 53 93
10.0 57 56 98
7.5 57 51 89
5.0 57 46 81
2.5 57 33 58
Probit analysis of the data determined that the concentration of HCV
RNA detected with 95% probability was 10.5 IU/mL A 9-member panel prepared by diluting HCV armored RNA from 8.33 log
(95% CI 8.6-14.0 IU/mL). IU/mL to 1.03 log IU/mL in HCV negative human serum was tested. The
results, representative of the Abbott RealTime HCV assay linearity, are
Limit of Detection, 0.2 mL Sample Volume shown in Figure 2.
The LOD of the Abbott RealTime HCV assay is 30 IU/mL with the 0.2 mL
sample preparation procedure and was determined as described for the Figure 2. Serum
0.5 mL sample preparation procedure. The results, representative of the
analytical sensitivity performance of the Abbott RealTime HCV assay, are
summarized in Table 2.
Table 2.
Number Number Percent
(IU/mL) Tested Detected Detected
50.0 60 59 98
35.0 60 60 100
25.0 60 59 98
20.0 59a 48 81
15.0 60 53 88
12.5 60 54 90
10.0 60 39 65
7.5 60 27 45
a One sample was inhibited and was deleted from the data analysis.
Probit analysis of the data determined that the concentration of HCV
RNA detected with 95% probability was 23.8 IU/mL
(95% CI 17.4-59.7 IU/mL).
Linear Range
The upper limit of quantitation (ULQ) for the Abbott RealTime HCV Linearity analysis was performed following the NCCLS EP6-A26 guideline.
assay is 100 million IU/mL and the lower limit of quantitation is The Abbott RealTime HCV assay was shown to be linear in serum and
equivalent to LOD (12 IU/mL for the 0.5 mL sample preparation plasma across the range of HCV RNA concentrations tested.
procedure, 30 IU/mL for the 0.2 mL sample preparation procedure). Precision
A 9-member panel prepared by diluting HCV armored RNA from 8.21 The precision of the Abbott RealTime HCV assay was evaluated for
log IU/mL to 0.91 log IU/mL in HCV negative human plasma was tested. the 0.5 mL sample preparation procedure using the Abbott m1000 and
The results, representative of the Abbott RealTime HCV assay linearity, the Abbott m2000sp instruments and the manual sample preparation
are shown in Figure 1. method. An 8-member HCV RNA panel was prepared. Panel members 1,
3, and 5 were dilutions of an HCV viral stock in negative human plasma.
Panel members 2 and 4 were dilutions of an HCV viral stock in negative
human serum. Panel members 6 through 8 were prepared by diluting
HCV armored RNA in negative human plasma. One lot of amplification
reagents was assigned to each of 3 Abbott m2000 instruments. For
the precision studies with the Abbott m1000 and the Abbott m2000sp
the panel members were tested in replicates of 4 in the first run on
each instrument pair and replicates of 5 in each subsequent run, for
a total of 15 runs. For the precision study using the manual sample
preparation method, new dilutions of panel members 1, 2, 3, 5, 7,
and 8 were prepared. These panel members were tested in replicates
of 2 or 3 in 5 runs on each Abbott m2000rt instrument. Analysis was
performed according to NCCLS EP10-A2.27 Within-run, between-run,
and inter-assay (within-run and between-run) standard deviations were
determined. The Abbott RealTime HCV assay was designed to achieve
an inter-assay standard deviation (SD) of less than or equal to 0.25 log
IU/mL of HCV RNA for samples containing HCV concentrations from
100 to 100 million IU/mL. The results, representative of the precision of
the Abbott RealTime HCV assay, are summarized in Tables 3, 4, and 5.
9
 Table 3. Precision with the Abbott m1000 and Abbott Drug Pool Drugs Tested
m2000rt Instruments 1 Zidovudine, Saquinavir, Ritonavir, Clarithromycin,
Conc. Within‑Run Between‑Run Interferon 2a, Interferon 2b
Panel Mean Conc. Mean Component Component Inter-Assay 2 Abacavir sulfate, Amprenavir, Peginterferon 2a,
Member n (IU/mL) (Log IU/mL) SDa SDa SDa,b Peginterferon 2b, Ribavirin
1 66c,d 11 1.03 0.26 0.00 0.26 3 Tenofovir disoproxil fumarate, Lamivudine, Indinavir
2 72 92 1.96 0.08 0.07 0.11 sulfate, Ganciclovir, Valganciclovir hydrochloride,
3 72 664 2.82 0.04 0.07 0.09 Acyclovir
4 72 9,182 3.96 0.05 0.05 0.07 4 Stavudine, Efavirenz, Lopinavir, Enfuvirtide, Ciprofloxacin
5 70d,e 62,682 4.80 0.05 0.07 0.08 5 Zalcitabine, Nevirapine, Nelfinavir, Azithromycin,
6 71e 917,350 5.96 0.05 0.07 0.08 Valacyclovir
7 72 10,599,314 7.03 0.07 0.08 0.11 The assay was also evaluated by testing 70 specimens that had
8 71d 113,005,242 8.05 0.03 0.06 0.07 been either obtained from individuals diagnosed or screened for an
a
autoimmune disorder (systemic lupus erythematosus [SLE], anti‑nuclear
Standard deviations (SD) are in log IU/mL. antibodies [ANA], and rheumatoid factor [RF]) or serologically
b Inter-assay contains within-run and between-run components. characterized as positive for the following viral markers: HBsAg, anti-
c HCV RNA was not detected in 5 replicates. HTLV-I/II, anti-HIV-1, anti-HIV-2. An additional 10 specimens that were
d One replicate was deleted from the data analysis due to an instrument error. PCR positive for a flavivirus (West Nile virus [n=4] and GB virus C [n=6])
e One replicate was aborted during sample preparation due to an instrument error. were tested.
Table 4. Precision with the Abbott m2000sp and Abbott HCV RNA was not detected in 77 of the 80 specimens tested. HCV RNA
m2000rt Instruments was detected, but less than LOD, in 3 specimens (1 RF, 1 SLE and 1
anti-HIV-1). Insufficient volume did not allow for confirmation.
Conc. Conc. Within‑Run Between‑Run
Panel Mean Mean Component Component Inter-Assay Specificity
Member n (IU/mL) (Log IU/mL) SDa SDa SDa,b The target specificity of the Abbott RealTime HCV assay is greater than
1 65c 8 0.88 0.25 0.00 0.25 or equal to 99.5% after resolution. In a representative study, specificity
2 72 91 1.96 0.08 0.04 0.09 was evaluated by testing 56 HCV seronegative serum and 56 HCV
seronegative plasma specimens. The specimens were tested on 3
3 72 564 2.75 0.05 0.03 0.06
Abbott m2000 instrument systems with 3 lots of amplification reagents.
4 72 8,853 3.95 0.03 0.02 0.04 HCV RNA was not detected, resulting in 100% (112/112) specificity (95%
5 71d 61,320 4.79 0.04 0.02 0.04 CI 96.76-100.00%).
6 71d 925,118 5.97 0.04 0.02 0.04
Cross-Reactivity
7 71d 10,045,506 7.00 0.07 0.01 0.07
The following viruses and microorganisms were evaluated for potential
8 71d 109,257,360 8.04 0.04 0.01 0.04 cross-reactivity in the Abbott RealTime HCV assay. Purified nucleic acid
a Standard deviations (SD) are in Log IU/mL. or viral lysate from each microorganism or virus was added to HCV RNA
b Inter-assay contains within-run and between-run components. negative samples and samples that contained 1,000 IU/mL HCV RNA.
c HCV RNA was not detected in 7 replicates. Human immunodeficiency virus 1 Vaccinia virus
d One replicate was deleted from the data analysis due to an instrument error. Human immunodeficiency virus 2 BK human polyomavirus
Table 5. Precision with Manual Sample Preparation Method Human T-lymphotropic virus 1 Human papilloma virus 16
Within‑Run Between‑Run Hepatitis B virus Human papilloma virus 18
Panel Conc. Mean Conc. Mean Component Component Inter-Assay Epstein-Barr virus Neisseria gonorrhoeae
Member n (IU/mL) (Log IU/mL) SDa SDa SDa,b
Herpes simplex virus 1 Chlamydia trachomatis
1 41c 8 0.93 0.20 0.00 0.20
Herpes simplex virus 2 Candida albicans
2 42 79 1.90 0.09 0.03 0.10
Cytomegalovirus Staphylococcus aureus
3 42 651 2.81 0.06 0.08 0.10
Human herpesvirus 6B Staphylococcus epidermidis
5 41d 51,114 4.71 0.09 0.08 0.12
42 9,628,784 6.98 0.07 0.01 0.07
Human herpesvirus 8 Mycobacterium gordonae
7
8 42 95,761,398 7.98 0.05 0.04 0.06 Varicella-zoster virus Mycobacterium smegmatis
a
Dengue virus 4
Standard deviations (SD) are in log IU/mL.
b Inter-assay contains within-run and between-run components.
No interference in the performance of the Abbott RealTime HCV assay
c was observed in the presence of the potential cross-reactants for all
HCV RNA was not detected in 1 replicate.
d positive and negative samples tested.
One replicate was deleted from the data analysis due to an instrument error.
Detection and Quantitation of HCV Genotypes
Potentially Interfering Substance
The ability of the Abbott RealTime HCV assay to detect and quantitate
The susceptibility of the Abbott RealTime HCV assay to interference by
HCV genotypes was evaluated in 3 studies. In the first study, dilution
elevated levels of endogenous substances was evaluated. HCV negative
linearity was demonstrated by diluting 6 specimens, one of each
samples and samples containing 1,000 IU/mL of HCV RNA were tested.
genotype 1 through 6, to target concentrations of 5.0 log IU/mL, 3.5 log
No interference in the performance of the Abbott RealTime HCV assay IU/mL, 2.0 log IU/mL, and 1.0 log IU/mL. Four replicates were tested at
was observed in the presence of the following substances for all HCV each concentration of each genotype. These representative data are
positive and negative samples tested: shown in Figure 3. The correlation coefficients ranged from 0.994 to
• Hemoglobin 500 mg/dL 0.998.
• Triglycerides 3000 mg/dL
• Bilirubin 20 mg/dL
• Protein 9 g/dL
Antivirals and antibiotics at concentrations in excess of peak plasma or
serum levels were tested in 5 pools. No interference in the performance
of the Abbott RealTime HCV assay was observed in the presence of the
following drug pools for all HCV positive and negative samples tested:

10
Figure 3 Figure 4.

0
0 1 2 3 4 5 6 7

In the third study, purified RNA transcripts from genotypes 1 through 6

were diluted to a target concentration of 1,000 IU/mL and tested. The
results are summarized in Table 7. The results demonstrated equivalent
In the second study, a total of 75 specimens representative of each quantitation of the genotypes tested.
genotype were tested with the Abbott RealTime HCV assay. The Table 7.
results are summarized in Table 6. The correlation between the Abbott
HCV Mean Concentration
RealTime HCV assay results and those of a comparator assay is shown
Genotype (log IU/mL)
in Figure 4. Method comparison analysis was performed according to
NCCLS Document EP9-A2.28 1 3.00
2 3.04
Table 6.
3 2.98
HCV Number of Number of Percent
Genotypes Specimens Tested Specimens Detected Detected 4 2.80
1 14 14 100 5 2.82
1a 3 3 100 6 3.14
1b 12 12 100 Correlation
2 9 9 100 Specimens from 131 HCV infected patients were tested with the Abbott
2a/2c 1 1 100 RealTime HCV assay and a comparator assay. Method comparison
2b 2 2 100 analysis was performed following NCCLS EP9-A2.28 The correlation plot
is shown in Figure 5. One specimen was identified as an outlier and
3a 12 12 100 upon re‑analysis excluding the outlier, the correlation coefficient
3b 1 1 100 was 0.950.
3c 1 1 100
Figure 5.
4 9 9 100
4a 3 3 100
4c/4d 1 1 100
5a 4 4 100
6 3 3 100

11
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2002;347(13):975-982. For technical assistance, call Abbott Molecular Technical Services at
10. Hadziyannis SJ, Sette H Jr, Morgan TR, et al. Peginterferon-alpha +49-6122-580, email molecularsupport@abbott.com, or visit the Abbott
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treatment of hepatitis C. Hepatology. 2004;39(4):1147-1171. THE PURCHASE OF THIS PRODUCT ALLOWS THE PURCHASER TO
12. Gerken G, Rothaar T, Rumi MG, et al. Performance of the COBAS USE IT FOR AMPLIFICATION OF NUCLEIC ACID SEQUENCES AND
AMPLICOR HCV MONITOR test, version 2.0, an automated reverse FOR DETECTION OF NUCLEIC ACID SEQUENCES FOR HUMAN IN
transcription-PCR quantitative system for hepatitis C virus load VITRO DIAGNOSTICS. NO GENERAL PATENT OR OTHER LICENSE
determination. J Clin Microbiol. 2000;38(6):2210-2214. OF ANY KIND OTHER THAN THIS SPECIFIC RIGHT OF USE FROM
PURCHASE IS GRANTED HEREBY. THIS PROVISION DOES NOT
13. Germer JJ, Heimgartner PJ, Ilstrup DM, et al. Comparative evaluation
PROHIBIT THE RESALE OF THIS PRODUCT.
of the VERSANT HCV RNA 3.0, QUANTIPLEX HCV RNA 2.0,
and COBAS AMPLICOR HCV MONITOR version 2.0 assays for Armored RNA® is a patented technology jointly developed by Ambion,
quantification of hepatitis C virus RNA in serum. J Clin Microbiol. Inc. and Cenetron Diagnostics, LLC. US patents #5,677,124, #5,919,625,
2002;40(2):495-500. #5,939,262 and patents pending.
Armored RNA is a registered trademark of Ambion, Inc.
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of the LCx HCV RNA quantitative assay. J Virol Methods. ProClin is a registered trademark of Rohm and Haas.
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standard for nucleic acid amplification technology (NAT) assays for subsidiaries in the US and/or certain other countries.
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subsidiaries in the US and/or certain other countries.
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by a Thermus thermophilus DNA polymerase. Biochemistry. Abbott m, m1000, m2000, RealTime, m2000rt, and m2000sp are
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Abbott Molecular Inc. is the legal manufacturer of
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DC: US Government Printing Office; December 2009. Ring 2, 65205 Wiesbaden, Germany.
[Also available online. Type> www.cdc.gov, search>BMBL5>look up
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Geneva, Switzerland: World Health Organization; 2004. © 2005, 2020 Abbott. All Rights Reserved.
www.abbottmolecular.com
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June 2020
hepatitis C virus RNA with a solid-phase signal amplification method:
definition of optimal conditions for specimen collection and clinical 51-602124/R12
application in interferon-treated patients. Hepatology. 1994(6);
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