Verification and application of the HAV antigen detection ELISA kit

Objective To verify the consistency of the ELISA kit prepared in original manufacturing site for the HAV antigen detection after manufacturing site change of inactivated hepatitis A vaccine (human diploid cell). Methods The ELISA kit was prepared with the anti-HAV polyclonal antibody-coated plate and the horseradish peroxidase-labeled anti-HAV antibody prepared in original manufacturing site, and its linearity, accuracy, precision, specificity and stability were verified. Meanwhile, ELISA kit was used to detect HAV antigen concentrations in the intermediate products of inactivated hepatitis A vaccine. Results In new manufacturing site, the linear range of the ELISA kit was 0.2-6.4 IU/ml, and the linear determination coefficient was >0.99.The precision of the ELISA kit was good, and the coefficients of variation were all <15%. The recovery rates of samples with different HAV antigen concentrations were 84%-104%. The ELISA kit specifically detected HAV antigen, and did not react with non-HAV and cell matrix for HAV culture. There was no effect on the detection results when the antibody-coated plate was placed at 37 ℃ for 7 d. The detection results showed that the average antigen contents of HAV harvests, HAV purified liquid and final bulk of vaccine were 213.25, 146.70 and 48.91 IU/ml, respectively, meeting with decline pattern of the HAV antigen content in the process. Conclusion The ELISA kit prepared in original manufacturing site can be used to detect the inactivated hepatitis A vaccine (human diploid cell) prepared in new manufacturing site. Key words: Hepatitis A antigens; Hepatitis A vaccines; Enzyme-linked immunosorbent assay