OnSite HCV Ab Plus Rapid Test - Cassette (Serum / Plasma) Page 1 of 2

waste.
11. Handle the negative and positive controls in the same manner as patient specimens.
12. The test result should be read 15 minutes after a specimen is applied to the sample well
or sample pad of the device. Reading the result after 20 minutes may give erroneous
Catalog Number R0
R0023C results.
13. Do not perform the test in a room with strong air flow, i.e.
i an electric fan or strong air-
conditioning.

REAGENT PREPARATION AND STORAGE INSTRUCTIONS

In vitro Diagnostic
INTENDED USE All reagents are ready to use as supplied. Store unused test devices
device unopened at 2-30°C. The
positive and negative controls should be kept at 2--8°C or the temperature indicated. If stored at
The OnSite HCV Ab Plus Rapid Test is a double antigen lateral flow chromatographic 2-8°C,, ensure that the test device is brought to room temperature before opening. The test
immunoassay for the qualitative detection of anti-hepatitis C virus antibodies (IgG, IgM, IgA) in device is stable through the expiration date printed on the sealed pouch. Do not freeze the kit
human serum or plasma. It is intended to be used as a screening test and as an aid in the or expose the kit to temperatures over 30°C.
diagnosis of infection with HCV. Any reactive specimen with the OnSite HCV Ab Plus Rapid
Test must be confirmed with alternative testing method(s) and clinical findings. SPECIMEN COLLECTION AND HANDLING

SUMMARY AND EXPLANATION OF THE TEST Consider any materials of human origin as infectious and handle them using standard bio-
bio
safety procedures.
Hepatitis C virus (HCV), which was formerly described as the parentally transmitted form of
1 2
non-A, non-B hepatitis (NANBH) , causes chronic disease in 50% of patients . HCV can also Plasma
be transmitted through intravenous drug abuse and sexual contact3. Step 1: Collect blood specimen into a lavender, blue or green top collection tube (containing
EDTA, citrate or heparin, respectively in Vacutainer® ) by venipuncture.
Hepatitis C virus is a single-stranded RNA virus with structural similarities to the flavivirus Step 2: Separate the plasma by centrifugation.
family. Nucleic acid sequences of HCV cDNA clones provide the basis for the construction of Step 3: Carefully withdraw the plasma into a new pre-labeled
pre tube.
recombinant peptides representing putative hepatitis C virus proteins4,5. Anti
Anti-hepatitis C virus
antibody screening of blood using synthetic or recombinant proteins helped to identify Serum
apparently healthy blood donors with anti-HCV HCV antibodies who otherwise might have Step 1: Collect blood specimen into a red top collection tube (containing no anticoagulants
anticoag in
transmitted the virus6. Therefore, the OnSite HCV Ab Plus Rapid Test is a useful tool for blood Vacutainer®) by venipuncture.
bank screening safety. Step 2: Allow the blood to clot.
Step 3: Separate the serum by centrifugation.
The OnSite HCV Ab Plus Rapid Test was developed to detect anti-HCV
HCV antibodies (IgG, IIgM, Step 4: Carefully withdraw the serum into a new pre-labeled
pre tube.
IgA) in human serum or plasma. The test can be performed by minimally trained personnel and
without cumbersome laboratory equipment. Test specimens as soon as possible after collecting. Store specimens at 2-8°C, if not tested
immediately. Specimens can be stored at 2-8°C for up to 5 days. The specimens should be
TEST PRINCIPLE frozen at -20°C for longer storage.
The OnSite HCV Ab Plus Rapid Test is a double antigen lateral flow chromatographic Avoid multiple freeze-thawthaw cycles. Prior to testing, bring frozen specimens to room
immunoassay. The test cassette consists of: 1) a burgundy colored conjugate pad containing temperature slowly and mix gently. Specimens containing visible particulate matter should be
recombinant HCV fusion antigen (core, NS3, NS4 and NS5) conjugated with colloid colloidal gold clarified by centrifugation before testing.
(HCV Ag conjugates) and a control antibody conjugated with colloidal gold
gold, 2) a nitrocellulose
membrane strip containing a test line (T line) and a control line (C line
line). The T line is pre- Do not use samples demonstrating gross lipemia,
mia, gross hemolysis or turbidity in order to avoid
coated with recombinant HCV fusion antigen (core, NS3, NS4 and NS5) NS5), and C line is pre- interference with result interpretation.
coated with a control line antibody.
ASSAY
SAY PROCEDURE

Step 1: Bring the specimen and test components to room temperature

tempera if refrigerated or
frozen. Mix the specimen well prior to assay once thawed.

Step 2: When ready to test, open the pouch at the notch and remove the device. Place the
test device on a clean, flat surface.

Step 3: Be sure to label the device with the specimen’s ID number.

Step 4: Fill in the plastic dropper with the specimen.

specimen
When an adequate volume of test specimen is dispensed into the sample well of the test
cassette, the specimen migrates by capillary action acrosss the cassette. Antibodies to HCV, if Holding the dropper vertically, dispense 1 drop (about 30-45 µL) of specimen into
present in the specimen, will bind to the HCV Ag conjugates. The immunocomplex is then the center of the sample well making sure that there are no air bubbles.
captured on the membrane by the pre-coated, non-conjugated HCV fusion antigen forming a
burgundy colored T line, indicating a HCV Ab positive or reactive test result. Absence of the T Immediately add 1 drop (about 35-50 µL)
µL of sample diluent to the sample well with
line suggests a negative result. the bottle positioned vertically.

The test contains an internal control (C line), which should exhibit a burgundy colored line of
the immunocomplex of control antibodies regardless of color development on the T line. If the
C line does not develop, the test result is invalid, and the specimen must be retested with Result
another device.
15 minutes
REAGENTS AND MATERIALS PROVIDED

1. Individually sealed foil pouches containing:

a. One cassette device 1 drop of specimen 1 drop of sample diluent
b. One desiccant
2. Plastic droppers Step 5: Set up a timer.
3. Sample Diluent (REF SB-R0023, 5 mL/bottle)
4. One package insert (instruction for use) Step 6: Read the result in 15 minutes. Positive results
result may be visible in as soon as 1
minute.
MATERIALS REQUIRED BUT NOT PROVIDED
Do not read result after 20 minutes. To avoid confusion, discard the test device
devic after
1. Clock or Timer
interpreting the result.
WARNINGS AND PRECAUTIONS QUALITY CONTROL
For in Vitro Diagnostic Use 1. Internal Control: This test contains a built-in
built control feature, the C line. The C line
1. This package insert must be read completely before performing the test. Failure to develops after adding the specimen and the sample diluent.
diluent If the C line does not
follow the insert may lead to inaccurate test results. develop, review the entire procedure and repeat the test with a new device.
2. Do not open the sealed pouch unless ready to conduct the assay.
3. Do not use expired devices. 2. External Control: Good Laboratory Practice recommends using external controls,
4. Bring all test materials to room temperature (15-30°C) before use. positive and negative, to ensure the proper performance of the assay, particularly
particul under
5. Do not use the components in any other type of test kit as a substitute for the the following circumstances:
components in this kit. a. A newew operator uses the kit, prior to performing the testing of specimens.
6. Do not use hemolized blood specimens for testing. b. A new lot of test kits is used.
7. Wear protective clothing and disposable gloves while handling the kit reagents and c. A new shipment of kits is used.
clinical specimens. Wash hands thoroughly after performing the test. d. The temperature used during storage of the kits kit fall outside of 2-30°C.
8. Users of this test should follow the US CDC Universal Precautions for prevention of e. The temperature of the e test area falls outside of 15-30°C.
15
transmission of HIV, HBV and other blood-borne pathogens. f. To verify a higher than expected frequency of positive or negative results.
results
9. Do not smoke, drink or eat in areas where specimens or kit reagents are being handled. g. To investigate the cause of repeated invalid results.
results
10. Dispose of all specimens and materials used to perform the test as bio bio-hazardous
 OnSite HCV Ab Plus Rapid Test - Cassette (Serum / Plasma) Page 2 of 2

INTERPRETATION OF ASSAY RESULT

LIMITATIONS OF TEST
1. NEGATIVE RESULT: If only the C line is developed, the test indicates that no
detectable antibodies to HCV are present in the specimen. The result is negative or 1. The Assay Procedure and the Interpretation of Assay Result sections must be followed
non-reactive. closely when testing for the presence of antibodies to HCV in serum or plasma from
individual subjects. Failure to follow the procedure may lead to inaccurate results.
2. The OnSite HCV Ab Plus Rapid Test is limited to the qualitative detection of antibodies
anti-HCV in human serum or plasma. The intensity of the test line does not have linear
correlation with the antibody titer in the specimen.
3. A non-reactive result for an individual subject indicates the absence of detectable
antibodies to HCV. However, a non-reactive test result does not preclude the possibility
2. POSITIVE RESULT: If both the C and T lines are developed, the test indicates the of exposure to or infection with HCV.
presence of antibodies to HCV in the specimen. The result is positive or reactive. 4. A non-reactive result can occur if the quantity of antibodies to HCV present in the
specimen is below the detection limits of the assay or if the antibodies that are detected
are not present during the stage of disease in which a sample is collected.
5. Some specimens containing unusually high titers of heterophile antibodies or
rheumatoid factor may affect expected results.
6. If symptoms persist when the result from OnSite HCV Ab Plus Rapid Test is non-
reactive, it is recommended to re-sample the patient a few days later or test with an
Samples with reactive results should be confirmed with alternative testing method(s)
alternative test device.
and clinical findings before a positive determination is made.
7. Results obtained with this test should only be interpreted in conjunction with other
3. INVALID: If no C line is developed, the assay is invalid regardless of color development diagnostic procedures and clinical findings.
on the T line as indicated below. Repeat the assay with a new device.
REFERENCES

1. Kuo,G, Choo Q-L, Alter, HJ, et al. An assay for circulating antibodies to a major
etiologic virus of human non-A, non-B hepatitis. Science 1989. 244:362-4.
2. Esteban JI, Gonzalez A, Hernandez JM, et al. Evaluation of antibodies to hepatitis C
virus in a study of transfusion-associated hepatitis. N Engl J Med 1990. 323:1107-12.
3. Miyamura T, Saito I, Katayama T, et al. Detection of antibody against antigen
PERFORMANCE CHARACTERISTICS expressed by molecularly cloned hepatitis C virus cDNA: application to diagnosis and
blood screening for posttransfusion hepatitis. Proc Natl Acad Sci USA 1990. 87:983-7.
1. Clinical Performance 4. Estaban JI, Esteban R, Viladomiu L, et al. Hepatitis C virus antibodies among risk
A total of 1050 samples from susceptible subjects were tested with the OnSite HCV Ab groups in Spain. Lancet 1989. 2:294-7.
Plus Rapid Test and with a commercial HCV ELISA kit. Comparison of the results for all 5. Houghton M, Weiner A, Han J, Kuo G, Choo Q-L. Molecular Biology of the Hepatitis C
subjects is shown in the following table. viruses: Implications for diagnosis, Development, and Control of Viral Disease.
Hepatology 1991. 14:381-8.
OnSite HCV Ab Plus Rapid Test 6. Alter HJ, Purcell RH, Shih JW, Melpolder JC, Houghton M, Choo Q-L, Kuo G. Detection
HCV ELISA Positive Negative Total of antibody to hepatitis C virus in prospectivity followed transfusion recipients with acute
Positive 312 4 316 and chronic non-A,non-B hepatitis. N Engl J Med 1989. 321:1494-1500.
Negative 3 731 734
Index of Symbols
Total 315 735 1050
Consult For in vitro
Relative Sensitivity: 98.7%, Relative Specificity: 99.6%, Overall Agreement: 99.3% instructions for use diagnostic use only Use by

2. Worldwide Performance Panel Tests per kit

REF Catalog # Lot Number
BBI's (Boston Biomedica Inc.) worldwide performance panel (WWHV301) was tested N
with the OnSite HCV Ab Plus Rapid Test. The results are shown in the following table. Store between 2-30°C Do not reuse
OnSite HCV Ab
Member ID Origin Genotype Abbott EIA
Plus Rapid Test Manufacturer Date of manufacture
301-01 Argentina 1b Positive Positive
301-02 Argentina 1b Positive Positive
301-03 Argentina 3a/b Positive Positive
301-04 Argentina 2a/c Positive Positive
CTK Biotech, Inc.
301-05 Argentina Not tested Negative Negative
10110 Mesa Rim Road
301-06 Uganda 4c/d Positive Positive San Diego, CA 92121, USA
301-07 Uganda Not tested Positive Positive Tel: 858-457-8698
301-08 Ghana Not tested Negative Negative Fax: 858-535-1739
301-09 China 1b, 2a/c Positive Positive E-mail: [email protected]
301-10 China 2 Positive Positive PI-R0023C Rev. E
301-11 China 1b Positive Positive Effective date: 2014-03-10
301-12 China 2 Positive Positive English version
301-13 China 1a/b, 2a/c Positive Positive For Export Only, Not For Resale In The USA
301-14 Egypt 3a Positive Positive
301-15 Egypt 4 Positive Positive
301-16 Egypt 4h Positive Positive
301-17 Egypt Not tested Positive Positive
301-18 USA 1b Positive Positive
301-19 USA 1a Positive Positive
301-20 USA 1a Positive Positive

3. Seroconversion Panel
BBI's (Boston Biomedica Inc.) seroconversion panel (PHV910 – (M)) was tested with
the OnSite HCV Ab Plus Rapid Test. The results are shown in the following table.

Abbott HCV EIA OnSite HCV Ab Plus

Member ID Days Bleeding
2.0 s/co* Rapid Test
910-01 0 0.2 Negative
910-02 4 0.3 Negative
910-03 8 1.3 Positive
910-04 11 2.9 Positive
910-05 15 2.4 Positive
* EIA results expressed as specimen absorbance to cut-off ratio (S/CO). Ratios > 1.0
are considered reactive.