Development and verification of an ELISA for quantitatively detecting Japanese encephalitis virus antigen / 临床检验杂志

ABSTRACT

Objective To develop a double antibody sandwich ELISA for quantitatively detecting Japanese encephalitis virus(JEV) antigen.Methods The anti-JEV polyclonal antibodies were used to coat ELISA plates.Anti-JEV monoclonal antibodies were used as enzyme-labeled conjugate.A standard curve based on known amounts of JEP antigen was established by the ELISA.Various parameters of the assay were analyzed.Results The optimal linear range was 12.5～200 U/ml(r=0.9989).The quantitation limit was 12.5 U/ml.The recovery rate for the accuracy test was 85.0%～103.3%.The coefficients of variation for intra-assay and inter-assay precision were 4.3%and 5.5%respectively.No cross-reaction was observed with HAV vaccine,influenza vaccine,Vero cell Iysates,newborn bovine serum,or human albumin.Conclusions The data indicate that the ELISA developed in this study has high specificity,precision, accuracy,and stability.The assay should be suitable for quantitative determination of JEV antigen in various vaccine products.