LIBEN PRIMARY HOSPITAL

LABORATORY STANDARD
OPERATING PROCEDURE
FOR
HEPATITS C VIRUS

Compiler: Mekuant&Getachew Signature:

Approver: Abere abye Signature:

Effective Date: Sep. 2013 Revision No. 0

Document No:LPHL/SOP5.5-004 Copy No.

.
I. REVISION AND AMENDMENT
A. Annual Review of Document
Reviewed by: Approved by:
Revision No. Review Date
Name Signature Name Signature

B. Version Change History/Description

Version Effectiv Description of Version Change Name & Name &

. No. e Date Signature of Signature of

NOTE: This is a CONTROLLED document for internal use only.

 LIBEN PRIMARY HOSPITAL LABORATORYSTANDARD OPERATING Document No:
PROCEDURE LPHL/ALS/SOP
Ver. No: 0
Page No: Effective date:
Sop for HCV Ag Test 2 of 8 29 Sep, 2018

Reviewer approval

C. Amendment
Rev. Page Description of Amendment Amend Effective Name &
No. No ment Date Signature of
Date approval

NOTE: This is a CONTROLLED document for internal use only.

 LIBEN PRIMARY HOSPITAL LABORATORYSTANDARD OPERATING Document No:
PROCEDURE LPHL/ALS/SOP
Ver. No: 0
Page No: Effective date:
Sop for HCV Ag Test 3 of 8 29 Sep, 2018

Purpose For the qualitative detection of antibodies to hepatitis C virusin human whole
blood ,serum or plasma

Abbreviations
HVC= hepatitis C virus
HBV= hepatitis B virus
HIV= human immunodeficiency virus

Materials Reagents
1. Disposable gloves
2. HBSag antibody test cassettes
3. Sample diluents ( 4ml) if applicable
4.Test tube

Supplies
1. Centrifuge
2. Test tube ruck

3. timer

Sample Sample type Amount required Transport and Stability

NOTE: This is a CONTROLLED document for internal use only.

 LIBEN PRIMARY HOSPITAL LABORATORYSTANDARD OPERATING Document No:
PROCEDURE LPHL/ALS/SOP
Ver. No: 0
Page No: Effective date:
Sop for HCV Ag Test 4 of 8 29 Sep, 2018

Storage
Serum, plasma, o.2-2ml stored at 2-8 stored at 2-8 degree
and whole blood degree centigrade centigrade if the test
if the test is to be is to be run within
run within one one week
week

Safety
Precautions
1. This package insert must be read completely before performing the test. Failure to
follow the insert gives inaccurate test results.
2. Do not open the sealed pouch, unless ready to conduct the assay.
3. Do not use expired devices.
4. Bring all reagents to room temperature (15°C-30°C) before use.
5. Do not use the components in any other type of test kit as a substitute for the
components in this kit.
6. Do not use hemolized blood specimen for testing.
7. Wear protective clothing and disposable gloves while handling the kit reagents and
clinical specimens. Wash hands thoroughly after performing the test.
8. Users of this test should follow the US CDC Universal Precautions for prevention
of transmission of HIV, HBV and other blood-borne pathogens.
9. Do not smoke, drink, or eat in areas where specimens are being handled.
10. Dispose of all specimens and materials used to perform the test as bio hazardous
waste.
11. Handle the Negative and Positive Control in the same manner as patient
specimens.

NOTE: This is a CONTROLLED document for internal use only.

 LIBEN PRIMARY HOSPITAL LABORATORYSTANDARD OPERATING Document No:
PROCEDURE LPHL/ALS/SOP
Ver. No: 0
Page No: Effective date:
Sop for HCV Ag Test 5 of 8 29 Sep, 2018

12. The testing results should be read within 15 minutes after a specimen is applied to
the sample well or sample pad of the device. Read result after 15 minutes may
give erroneous results.
13. Do not perform the test in a room with strong air flow, ie. an electric fan or strong
air-conditioning.
.

Procedure Step Action

1 1. When you are ready to begin testing, open the sealed pouch by tearing
along the notch. Remove the test kit from the pouch and use it as soon as
possible.
2 2. Following the illustration, dip the test strip with the arrow side pointing
down into the vessel of serum for about 10 seconds. Do not immerse past
the marker line. Take the strip out and lay it flat on a clean, dry and non-
absorbent surface
3 . Wait for 10 minutes and read results. It is important that the background is
clear before the result is read. Do not read results after 30 minutes.

Result Negative: Only one color band appears on the control (C) region. No apparent

NOTE: This is a CONTROLLED document for internal use only.

 LIBEN PRIMARY HOSPITAL LABORATORYSTANDARD OPERATING Document No:
PROCEDURE LPHL/ALS/SOP
Ver. No: 0
Page No: Effective date:
Sop for HCV Ag Test 6 of 8 29 Sep, 2018

Interpretation band on the test (T) region.

Positive: In addition to a pink colored control (C) band, a distinct pink colored
band will also appear in the test (T) region.
Invalid: A total absence of color in both (C) and (T) regions and no colored band
appears on the control (C) region is an indication of procedure error and/or
the test reagent has deteriorated. Repeat with a new test kit. If the problem
persists,discontinue using the test kit immediately and contact your local
distributor.Mark Line

LIMITATIONS
1. This test should be used for the detection of antibodies to HCV in serum samples.
2. Only detect the presence of Anti-HCV, it should not be used as the sole criteria
for the diagnosis of Hepatitis C viral infection.
3. As with all diagnostic tests, all results must be considered with other clinical
information available to the physician. A definite clinical diagnosis should only be
made by the physician after all clinical and laboratory findings have been evaluated.
4. If the test result is negative and clinical symptoms persist, additional follow-up
testing using other clinical methods is recommended. A negative result any time
does not preclude the possibility of Hepatitis C Virus infection.

Principle
One Step ANTI-HCV Test is based on the principle of double antigen sandwich
immunoassay for determination of anti-HCV in serum. Purified recombinant antigens
are employed to identify anti-HCV specifically. This one step test is very sensitive
and only takes 10-20 minutes for the result to be read. Test results are read visually
without any instrument.

NOTE: This is a CONTROLLED document for internal use only.

 LIBEN PRIMARY HOSPITAL LABORATORYSTANDARD OPERATING Document No:
PROCEDURE LPHL/ALS/SOP
Ver. No: 0
Page No: Effective date:
Sop for HCV Ag Test 7 of 8 29 Sep, 2018

Clinical Utility Hepatitis C virus (HCV) infection is a common cause of liver disease in thalassemia
major patients. The relationships of the infection to blood transfusion and the
infection's effects on liver function have also been determined

Reference 1. Engvall E, Perimann P. Enzyme-Linked Immunosorbent Assay (ELISA)

Quantitative Assay of Immunoglobulin G. Immunochem 1971:8:871-4.
2 . Engvall E.Perimann P. Enzyme-Linked Immunosorbent Assay (ELISA). In:
Pecters H, editor. Protides of the Biological Fluids. Proceedings of the
Nineteenth Colloquium, Burgge. Wxford: Pergamon Press, 1971:553-6.
3. Engvall E, lonsson K, Perimann P Enzyme-Linked mmunogolbulin G By
Means of Enzyme-Labelled Antigen and Antibody-Coated Tubes. Biochem
Biophys Acta 1971,
4. BanWeemen BK, Schuurs AHWM. Immunoassay Using Antigen-Enzyme
Conjugates. FEBS Letters 1971:15:232- 6.
5 Wisdom GB. Enzyme-Immunoassay.Clin Chem 1976;22:1243-55.
6 .Wolters G, Kuijpers L, Kacaki 1, Schuurs A. Solid-Phase

. Declaration
.

NOTE: This is a CONTROLLED document for internal use only.

 LIBEN PRIMARY HOSPITAL LABORATORYSTANDARD OPERATING Document No:
PROCEDURE LPHL/ALS/SOP
Ver. No: 0
Page No: Effective date:
Sop for HCV Ag Test 8 of 8 29 Sep, 2018

WE, the undersigned laboratory personnel, certify that we conducting every steps of the procedures
incorporated in this SOP after a prior reading.

Name Signature and Date

1. Abere Abye -----------------------------------------

2. Getachew Abebe -----------------------------------------

3. Mekuant Alemkere -----------------------------------------

4. Girma Zerefaw -----------------------------------------

NOTE: This is a CONTROLLED document for internal use only.