Initial Establishment Of Double Antigen Sandwich Detection Method For Hepatitis C Virus Antibodies

Hepatitis c virus (HCV) infection caused by hepatitis c is now a global distribution, there are nearly180million people of hepatitis c infection, and accounts for about3%of the world’s population, theinfection rate of about3.2%of our country.Currently used in the main of the third generation of indirect ELISA detection method because of itsinherent flaws in methodology, easy to cause false positive test results and targets; Development of doubleantigen sandwich ELISA method to detect reagent is the developing direction of the reagent, due to therelatively small molecular weight HCV antigen, using enzymes to HCV antigen directly, easy to cause thespace steric hindrance, make the combination of antibody and antigen further by inhibition of larger, so theHCV antigen markup is built against hepatitis c virus antibody of the main difficulties with double antigensandwich ELISA assay, is also the research hotspot in this field at home and abroad.This study through the amplification effect of biotin avidin system to reduce the space steric hindranceantigen tags, establishing anti HCV antibody double antigen sandwich ELISA detection method. Usinglaboratory has built plasmid vector to express and purify antigen sandwich ELISA method, the coupling tothe activation antigen on the long arm of biotin, preparation of growth arm biotinylated HCV antigen, as asandwich antigen, with horseradish peroxidase mildew affinity tag chain element as enzymes, usinghorseradish peroxidase catalytic chromogenic substrate, establish a double antigen sandwich method.The use of set up double antigen clamp method to detect500clinical specimens, and parallel tests, andother reagents on the test results are inconsistent with HCV RNA quantitative detection kit sampleverification. The results show that the detection results consistent with493samples, including48positivesamples and445negative samples; there are7inconsistent test results of samples including indirectpositive and clip art negative has4copies, test results for HCV RNA are negative; Clip art positive andnegative three of the indirect method, there are two test positive for HCV RNA.Through the data statistics, this study established the double antigen specificity of center method andindirect method were99.78%,98.89%, sensitivity100%,96%respectively. Indicates that the research ofdouble clip art the sensitivity and specificity of antigen are better than the indirect method, deserves further research.