procedure and follow the standard procedures for proper disposal of

specimens. Wear protective clothing such as laboratory coats, disposable

gloves and eye protection when specimens are assayed.
 Do not interchange or mix reagents from different lots. Do not mix solution
bottle caps.
HPV 16/18 Antigen  Humidity and temperature can adversely affect results.
 When the assay procedure is completed, dispose the swabs carefully after
Rapid Test Device autoclaving them at 121°C for at least 20 minutes. Alternatively, they can be
treated with 0.5% sodium hypochloride (or house-hold bleach) for one hour
500140 Specimen: Swab before disposal. The used testing materials should be discarded in
accordance with local, state and/or federal regulations.
Language: English Version: 02  Do not use cytology brushes with pregnant patients.
Effective Date: 2012-03
STORAGE AND STABILITY
For professional in vitro diagnostic use only.
 The kit should be stored at 2-30°C until the expiry date printed on the sealed
INTENDED USE pouch.
The StrongStep® HPV 16/18 Antigen Rapid Test Device is a rapid visual  The test must remain in the sealed pouch until use.
immunoassay for the qualitative presumptive detection of HPV 16/18 E6&E7  Do not freeze.
oncoproteins in female cervical swab specimens. This kit is intended to be used  Cares should be taken to protect components in this kit from contamination.
as an aid in the diagnosis of Cervical Pre-cancer and Cancer. Do not use if there is evidence of microbial contamination or precipitation.
Biological contamination of dispensing equipments, containers or reagents
INTRODUCTION can lead to false results.
In developing countries, cervical cancer is a leading cause of cancer related
death of women, due to the lack of implementation of screening tests for
SPECIMEN COLLECTION AND STORAGE
cervical pre-cancer and cancer. A screening test for low resource settings should  The quality of specimen obtained is of extreme importance. As much as
be simple, rapid, and cost effective. Ideally, such a test would be informative cervical epithelial cell should be collected by the swab.
regarding HPV oncogenic activity For cervical specimens:
Expression of both HPV E6 and E7 oncoproteins is essential for cervical cell  Use only Dacron or Rayon tipped sterile swabs with plastic shafts. It is
transformation to occur. Some research results demonstrated a correlation of E6 recommend to use the swab supplied by the kits manufacturer(The swab are
&E7 oncoprotein positivity with both severity of cervical histopathology and risk not contained in this kit, for the ordering information, please contact the
for progression. Hence, E6&E7 oncoprotein promises to be an appropriate manufacture or local distributor, the cataloge number is 207000). Swabs
biomarker of HPV-mediated oncogenic activity. from other suppliers have not been validated. Swabs with cotton tips or
PRINCIPLE wooden shafts are not recommended.
The StrongStep® HPV 16/18 Antigen Rapid Test Device has been designed to  Before specimen collection, remove excess mucus from the endocervical area
detect HPV 16/18 E6&E7 Oncoproteins through visual interpretation of color with a separate swab or cotton ball and discard. Insert the swab into the
development in the internal strip. The membrane was immobilized with monoclonal cervix until only the bottommost fibers are exposed. Firmly rotate the swab
anti-HPV 16/18 E6&E7 antibodies on the test region. During the test, the specimen is for 15-20 seconds in one direction. Pull the swab out carefully!.
allowed to react with colored monoclonal anti-HPV 16/18 E6&E7 antibodies colored  Do not place the swab in any transport device containing medium since
particals conjugates, which were precoated on the sample pad of the test. The mixture transport medium interferes with the assay and viability of the organisms is
then moves on the membrane by capillary action, and interact with reagents on the not required for the assay. Put the swab to the extraction tube, if the test
membrane. If there were enough HPV 16/18 E6&E7 oncoproteins in specimens, a may be run immediately. If immediate testing is not possible, the patient
colored band will form at the test region of the membrane. Presence of this colored samples should be placed in a dry transport tube for storage or transport. The
band indicates a positive result, while its absence indicates a negative result. swabs may be stored for 24 hours at room temperature (15-30°C) or 1 week
Appearance of a colored band at the control region serves as a procedural control. This at 4°C or no more than 6 month at -20°C. All specimens should be allowed
indicates that proper volume of specimen has been added and membrane wicking to reach a room temperature of 15-30°C before testing.
has occurred. PROCEDURE
KIT COMPONENTS Bring tests, specimens, buffer and/or controls to room temperature (15-
30°C) before use.
20 Individually packed test Each device contains a strip with colored
1. Prepare swab specimens:
devices conjugates and reactive reagents pre-
spreaded at the corresponding regions. • Place a clean extraction tube in the workstation. Add 8 drops of Extraction
1 bottle of Extraction Buffer A - Buffer solution containing 0.2 M sodium Buffer A to the extraction tube.
10ml hydroxide with yellow cap. • Immerse the patient swab into the extraction tube and wait 2 minutes. While
1 bottle of Extraction Buffer B - Buffer solution containing 0.2 M hydrochloric waiting, use a circular motion to roll the swab against the side of the
10ml acid with white cap. extraction tube so that the liquid is expressed from the swab and can
20 Extraction tubes For specimens preparation use. reabsorb.
1 Workstation Place for holding buffer vials and tubes. • At the end of the extraction time, add 8 drops of Extraction Buffer B to the
1 Package insert For operation instruction. tube and extract for another 1 minute in the same way. Then squeeze the
swab firmly against the tube to expel as much liquid as possible from the
MATERIALS REQUIRED BUT NOT PROVIDED swab. Discard the swab following guidelines for handling infectious agents.
• The extracted specimen can remain at room temperature for 60 minutes
Timer For timing use. without affecting the test result.
2. Remove the test from its sealed pouch, and place it on a clean, level surface.
PRECAUTIONS Label the device with patient or control identification. To obtain a best result,
the assay should be performed within one hour.
 For professional in vitro diagnostic use only. 3. Add 3 drops (approximately 100 µl) of extracted sample from the Extraction
 Do not use after expiration date indicated on the package. Do not use the Tube to the sample well on the test cassette.
test if its foil pouch is damaged. Do not reuse tests. Avoid trapping air bubbles in the specimen well (S), and do not drop any
 This kit contains products of animal origin. Certified knowledge of the origin solution in observation window.
and/or sanitary state of the animals does not totally guarantee the absence As the test begins to work, you will see color move across the membrane.
of transmissible pathogenic agents. It is therefore, recommended that these 4. Wait for the colored band(s) to appear. The result should be read at 15
products be treated as potentially infectious, and handled observing the minutes. Do not interpret the result after 20 minutes.
usual safety precautions (do not ingest or inhale).
 Avoid cross-contamination of specimens by using a new specimen collection INTERPRETATION OF RESULTS
container for each specimen obtained.
 Read the entire procedure carefully prior to performing any tests. POSITIVE RESULT: Two colored bands appear on the membrane. One
 Do not eat, drink or smoke in the area where the specimens and kits are band appears in the control region (C) and another
handled. Handle all specimens as if they contain infectious agents. Observe band appears in the test region (T).
established precautions against microbiological hazards throughout the
 1. Giovane C., et al., J. Mol. Recog. 1999;12:141-152.
C
2. Joo-Ho Kim, et al., Bull. Korean Chem. Soc. 2009;30;2999-3005
T
3. Sexually Transmitted Diseases, 4th Edition. King K. Holmes. McGraw-Hill
Professional, 2007.
NEGATIVE RESULT:
Only one colored band appears in the control region
C (C). No apparent colored band appears in the test GLOSSARY OF SYMBOLS
T region (T).
Catalog number Temperature limitation
INVALID RESULT: Control band fails to appear. Results from any test
which has not produced a control band at the Consult instructions for use Batch code
C C specified reading time must be discarded. Please In vitro diagnostic medical
T T review the procedure and repeat with a new test. If Use by
device
the problem persists, discontinue using the kit Contains sufficient for <n>
immediately and contact your local distributor. Manufacturer
tests
Authorized representative in
NOTE: Do not reuse
1. The intensity of the color in test region (T) may vary depending on the the European Community
concentration of aimed substances present in the specimen. But the CE marked according to IVD Medical Devices Directive 98/79/EC
substances level can not be determined by this qualitative test.
2. Insufficient specimen volume, incorrect operation procedure, or performing
expired tests are the most likely reasons for control band failure. Liming Bio-Products Co., Ltd,
No. 12 Huayuan Road,Nanjing, Jiangsu, 210042
QUALITY CONTROL P.R. China.
Tel: (0086)25 85476723 Fax: (0086)25 85476387
 Internal procedural controls are included in the test. A colored band
E-mail: sales@limingbio.com
appearing in the control region (C) is considered an internal positive
Website: www.limingbio.com
procedural control. It confirms sufficient specimen volume and correct
www.stddiagnostics.com
procedural technique.
www.stidiagnostics.com
 External controls are not supplied with this kit. It is recommended that positive
and negative controls be tested as a good laboratory practice to confirm the
WellKang Ltd.(www.CE-marking.eu) Tel: +44(20)79934346
test procedure and to verify proper test performance.
29 Harley St., London WIG 9QR,UK Fax: +44(20)76811874
LIMITATIONS OF THE TEST
1. The HPV 16/18 Anitgen Rapid Test Device (Swab) is for professional in vitro
diagnostic use, and should be used for the qualitative detection of HPV
16/18 E6&E7 Oncoproteins only. There is no meaning attributed to linen
color intensity or width.
2. Detection of HPV 16/18 E6&E7 Oncoproteins is dependent on the number of
proteins present in the specimen. This may be affected by specimen
collection methods and patient factors such as age, history of STD, presence
of symptoms, etc. The minimum detection level of this test may vary
according to serovar.
3. The expression of E6&E7 oncoprotein is only indicate the risk of cervical
cancer and pre-cancer occures, the positive results do not confirm the cancer
or pre-cancer occures and the negative results can not exclude the happening
of cervical cancer and pre-cancer.
4. As with all diagnostic tests, a confirmed diagnosis should only be made by a
physician after all clinical and laboratory findings have been evaluated.

PERFORMANCE CHARACTERISTICS

Table: StrongStep® HPV 16/18 E6&E7 Oncoproteins Test vs. IHC

IHC
Relative Sensitivity: 81.69%
+ - Total
Relative Specificity: 88.95%
StrongStep® + 58 21 79
Overall Agreement: 86.93% HPV Test
- 13 169 182

71 190 261

The antibody used in the HPV 16/18 Antigen test has been shown to
detect HPV type 16 and 18. Cross reactivity with other organisms has been
studied using suspensions of 10 7 org/ml. The following organisms were not
detected using the test:
Acinetobacter calcoaceticus Proteus vulgaris
Salmonella typhi Acinetobacter spp.
Staphylococcus aureus Candida albicans
Neisseria catarrhalis Neisseria gonorrhoea
Neisseria meningitidis Neiiseria lactamica
Escherichia coli Gardnerella vaginalis
Streptococcus faecalis Streptococcus faecium
Pseudomonas aeruginosa Trichomonas vaginalis
Ureaplasma Urealyticum Mycoplasma hominis

LITERATURE REFERENCES