Abstract: Objective To develop a sandwich ELISA for detection of mouse hepatitis virus (MHV). Method The ELISA was standardized by pair-matching experiment, and the working concentrations of the mAb 3D4H9 and HRP-conjugate mAb 4G9F2 were 12.8 ng/mL and 70.84 ng/mL, respectively, and judging with P/N>2 and absorbance_450nm ≥0.248 as positive criteria. Results The DAS-ELISA was specific for detection of MHV, but no cross-reaction with minute virus of mouse, reovirus type 3, sendai virus, pneumonia virus of mice. The intra-assay and inter-assay coefficient of variability were within 10%. In addition, a total 358 samples were tested by the DAS-ELISA, and indirect ELISA, there was no significant difference between results. Conclusion The DAS-ELISA was sensitive and specific which provided a useful tool for diagnosis of MHV.

Key words: Mouse hepatitis virus(MHV), Monoclonal antibodies, Double antibody sandwich ELISA