Specifications

Enzyme immunoassay for the detection of antibodies against Human Hepatitis Virus Type C (HCV)

HCV Ab ELISA Kit

 

Catalog No.:BE201A

 

Enzyme immunoassay for the detection of antibodies against Human Hepatitis Virus Type C (HCV)

                                                                                

INTRODUCTION

 

Hepatitis C virus (HCV), which was formerly described as the parent rally transmitted form of non-A, non-B hepatitis (NANBH)¹, becomes a chronic disease in 50% of the cases.²   HCV can also be transmitted through intravenous drug abuse, sexual , and household contact.³ Hepatitis C virus is a 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 proteins.^4,5Anti-hepatitis C virus antibody screening of blood using synthetic or recombinant proteins, helped to identify apparently healthy blood donors with anti-HCV antibodies who otherwise might have transmitted the virus.⁶

This is an enzyme linked immunosorbent assay using recombinant proteins derived from core regions of HCV virus to detect the presence of HCV antibodies in human sera.

 

 

Multiple epitomes’ of HCV proteins (Core, NS3, NS4 and NS5) are bound to the micro titer wells. When antibodies to HCV are present in the test sample, they react with recombinant proteins and attach to the solid-phase. Non-reactive antibodies are removed with the wash buffer. Human IgGs bound to the antigen are reacted with anti-human IgG peroxidase conjugate and visualized by subsequent reactions with a chromogenic substrate. Positive sample generates a medium to dark blue color. No color or very pale blue color indicates a negative reaction. The intensity of the reaction is photo metrically quantities.

 

 

Materials provided with the kits:

Item	Description	96T	480T
1	Coating Plate	1	5
2	Negative Control	1ml	5ml
3	Positive Control	1ml	5ml
4	Sample Diluents	12ml	60ml
5	Enzyme Conjugate	12ml	60ml
6	Wash Buffer Concentrate (20x)	30ml	150ml
7	Substrate Solution A	6ml	30ml
8	Substrate Solution B	6ml	30ml
9	Stop Solution	6ml	30ml
10	Plastic Bag	1	5
11	Seal Paper	3	15
12	Manual	1 copy	5 copy

 

Materials required but not provided:

1. Precision pipettes: 0.02, 0.05, 0.10, 0.15, 0.20, and 1.0 ml.
2. Disposable pipette tips.
3. Distilled water.
4. HumidifiedBox capable of maintaining 37°C
5. Absorbent paper or paper towel.
6. Micro titer plate or strip-well washer
7. Micro titer plate reader.

 

 

All human source material used in the preparation of this product was found to be negative for the presence of HCV antibodies, as well as for the hepatitis B surface antigen, using a commercial licensed method. Nevertheless, because no test method can offer complete assurance of the absence of infectious agents, this product should be handled with caution.

1. Avoid contact of reagents with the eyes and skin. If that occurs, wash thoroughly with water.
2. Wear gloves.
3. Do not pipette by mouth.
4. Do not smoke.
5. Dispose all used materials in a suitable biohazard us waste container. Remains of samples, controls, aspirated reagents and pipette tips should be collected in a container for this purpose and autoclaved 1-hour at 121°C or treated with 10% sodium hypochlorite (final concentration) for 30 min before disposal. (Remains containing acid must be neutralized prior addition of sodium hypochlorite).
6. Adjust washer to the plate used (flat bottom) in order to wash properly.
7. Do not mix reagents from different lots.
8. Do not use reagents after expiration date.
9. Extreme care should be taken to avoid microbial contamination and cross contamination of reagents.
10. Use a new pipette tip for each specimen and each reagent.
11. Soaps and/or oxidizing agents remaining in containers used for the substrate-TMB solution can interfere with the reaction.

 

 

Serum should be prepared from a whole blood specimen obtained by acceptable medical techniques. Either serum or plasma can be used in this test. Remove serum or plasma from the clot or blood cells as soon as possible to avoid hemolytic. Specimen with extensive particulate should be clarified by centrifugation prior to use. Specimen frozen at -20°C or colder may be used. Avoid repeated freeze thaw.

 

STORAGE OF TEST KIT

 

Unopened test kits should be stored at 2-8C upon receipt and the micro titer plate should be kept in a sealed bag to minimize exposure to damp air. Use up the reagents as soon as possible after the kit is unpacked.

 

 

I