动物造模,肝炎病毒 z-bio 2021-11-02 476

　　Objective To establish a mouse hepatitis virus (Mouse Hepatitis Virus, MHV) serum antibody ELISA detection method for daily monitoring of MHV in this area, so as to make timely judgments on the infection status of SPF mice.

　　Methods Three MHV strains MHV1, MHV-JHM and MHV-A59 were selected, purified and concentrated antigens were obtained by expansion and culture of L929 cells, and suitable types of coating antigens were selected; BALB/C mice were immunized to obtain high titer immunopositive serum; optimization The ELISA reaction system establishes a standardized ELISA kit and is used for the detection of clinical mouse serum samples.

　　Results The MHV coating antigen type suitable for the region was selected as MHV1, and a virus amplification and concentration purification method was established. The prepared MHV antiserum reached the level of a large number of high titer in the same batch and can be used as a standardized quality control serum. The best working concentrations of coated antigen, serum to be tested and enzyme conjugate are 4.0 μg/mL (10-7.73/0.1 mL TCID50), 1:40 and 1:4000 dilutions; average coefficient of variation within and between batches Respectively 5.13% and 5.57%; detection sensitivity is 1:4000 dilution; with mouse Sendai virus (SV), mouse pneumonia virus (PVM), reovirus type III (Reo3), mouse parvovirus (MVM) There is no cross-reaction with mousepox virus (Ect) positive sera. The relative deviation of the stability test is less than 10%. 165 serum samples were tested, and the matching rate of positive sera was 97.37% (37/38), and the matching rate of negative sera was 92.19% (118/127).

　　Conclusion The ELISA method established in this study has high specificity, sensitivity and reproducibility of results for the detection of MHV antibodies in mouse serum, and can be used for daily pathogenic monitoring of MHV in this area to make accurate judgments on the infection status of mice.