laboratory reagent Hepatitis C Virus test Antibody HCV ELISA test Kit

Specifications

1.EIA elisa test kit
2.in vitro diagnostic HCV
3.blood testing reagent
4.determine HCV elisa kit
5.blood chemistry test kit

Certificate

GMP

SFDA

ISO13485

ISO9001:2008

 

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.

 

 

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.

 

 

It is strongly advised to analyze each specimen and controls in duplicate. All the reagents should equilibrate to room temperature before use.

1. Dispense 100ml(or 3 drops) of specimen diluents into individualtest wells.
2. Dispense 100ml positive control and negative control duplicate into individual wells.
3. Add 10ml of each test sample into duplicate test wells; vortex to mix.
4. Incubate for 60 minutes at 37°C
5. Wash each well 5 times by filling each well with diluted wash buffer, then inverting the plate vigorously to get all water out and blocking the rim of wells on absorbent paper for a few seconds.
6. Add 100ml of Enzyme Conjugate to each well. Mix it gently by swirling the micro titer plate on flat bench for 1 minute. Do not add Enzyme Conjugate to the blank well.
7. Incubate for 30 minutes at 37°C
8. Wash the plate 5 times as step 5.
9. Add one drop (50ml) of Substrate Solution A (HRP-substrate) to each well, then add one drop (50ml) of Substrate Solution B (TMB) to each well. Mix gently and incubate at 37°C for 30 minutes. .
10. Add one drop (50ml) of Stop Solution to each well to stop the color reaction. Read the OD value at 450 nm/630 nm with dual filter plate reader. It is option to read the OD value at 450 nm with single filter plate reader. (using the OD value of the blank well to correct all the OD reading from all wells)