OneStep HCV #118770 Test

Atlas Link

InstaTest HCV

Rapid Hepatitis C Test

Cat. No. 118770-50 (50 tests)

introduction to InstaTest HCV

Hepatitis C virus (HCV), which was formerly described as the parentally transmitted form of non-A, non-B hepatitis (NANBH)1, becomes a chronic disease in 50% of the cases2. After transfusion from blood donors with HCV antibodies, 88% of recipients develop NANBH and seroconvert to HCV antibody positive2. HCV can also be transmitted through intravenous drug abuse, sexual and household contact3. Hepatitis C virus is single stranded RNA virus with some structural relations to the flavivirus family. Nucleic acid sequences of HCV cDNA clones provided the basis for the construction of recombinant peptides representing putative hepatitis C virus proteins4,5. Anti-hepatitis C virus antibody screening of blood donors with anti-HCV antibodies who otherwise might have transmitted the virus6.

We have developed a rapid immunofiltration assay using recombinant proteins derived from NS3 and core regions of HCV virus to detect the presence of HCV antibodies in human sera.

PRINCIPAL OF THE ASSAY

Recombinant proteins correspond to NS3 and core Hepatitis C are bound in the membrane of the filtration device. When antibodies to HCV are present in the test serum, they react with the recombinant proteins and attach to the solid-phase (next to T). A procedural control is included by attaching F(ab)’2 fragment of goat anti-human IgG (next to C). Non-reactive antibodies are filtered through with the wash buffer. Human antibodies Z(IgGs, IgMs and IgA) bound on the membrane are visualized by reacting with colloidal gold Protein A and Protein G. The appearance of TWO RED DOTS (one corresponding to the procedural control and one to the recombinant HCV proteins) indicates positive reaction. The appearance of a single red dot corresponding to procedural control indicates a negative reactions

COMPONENTS

  1. HCV filtration device with recombinant HCV proteins and procedural control.

2. Wash buffer in dropper vial

3. Colloidal gold conjugate in dropper vial

4. Sample dispenser - to add patient serum

STORE ALL REAGENTS AT 4C - 8C

PREPARATION FOR THE ASSAY

1. All reagents should always be brought to room

temperature, approximately 22C (72F), before use.

2. Allow time (around 30 seconds) for solution to be

totally absorbed before proceeding to next step.

TEST PROCEDURE

  1. Add 2 drops of wash buffer.
  2. Add 2 drops of patient serum. (Serum should be properly spinned down from the whole blood)
  1. Add 2 drops of wash buffer.
  2. Add 2 drops of colloidal gold conjugate.
  3. Add 2 drops of wash buffer.
  4. In positives, two red dots develop within 1-2 minutes. In negatives, one red dot will appear within 1-2 minutes.

RESULTS

Positive Result / Negative Result

Appearance of TWO RED MARKS in the filtration device indicates positive reactivity. Appearance of ONE RED MARK corresponding to “C” indicates negative reactivity. The color of the dot fades with time. OPTIONAL: Add 1 drop of fixative to the device to stabilize the developed dot. For good performance and sensitivity of the test, all devices and reagents should always be properly stored and transported, and, used in accordance with the expiration date.

Atlas Link, 12720 Dogwood Hills Lane, Fairfax, VA 22033 USA

Phone: (703) 266-5667, FAX: (703) 266-5664

OneStep HCV #118770 Test

OBSERVE UNIVERSAL PRECAUTIONS FOR HANDLING BODY FLUIDS. DECONTAMINATE AND DISPOSE OF WASTE PROPERLY.

REFERENCES:

  1. Kuo, G., Choo, Q.-L., Alter, H.J., et al. An essay for circulating antibodies to a major etiologic virus of human non-A, non-B hepatitis. Science 1989; 244:362-4.
  2. Esteban, J.I., Gonzalez, A., Hernandez, J.M., 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 expressed by molecularly loned hepatitis C virus cDNA: application to diagnosis and blood screening for posttransfusion hepatitis. Proc. Natl. Acad. Sci. USA 1990; 87:983-7.
  4. Esteban, J.I., Esteban, R., Viladomiu, L., et al. Hepatitis C virus antibodies among risk groups in Spain. Lancet 1989; 2:294-7.
  5. Choo, Q.-L., Juo, G., Weiner, A.J., Overby, L.R., Bradley, D.W. & Houghton, M. (1989) Isolation of a cDNA Clone Derived from Blood-borne Non-A, Non-B Viral Hepatitis Genome. Science 244, 359-362.
  6. Houghton, M., Weiner, A., Han, J., Kuo, G. & Choo, Q.-L. (1991) Molecular Biology of the Hepatitis C Viruses: Implications for Diagnosis, Development and Control of Viral Disease. Hepatology 14, 381-388.
  7. Alter, H.J., Purcell, R.H., Shih, J.W., Melpolder, J.C., Houghton, M., Choo, Q.-L. & Kuo, G. (1989) Detection of antibody to hepatitis C virus in prospectively followed transfusion recipients with acute and chronic non-A, non-B hepatitis. N. Engl. J. Med. 321, 1494-1500.

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Rev. 12/97

Atlas Link, 12720 Dogwood Hills Lane, Fairfax, VA 22033 USA

Phone: (703) 266-5667, FAX: (703) 266-5664