动物造模,肝炎病毒,血清抗体 z-bo 2020-09-25 367

　　Objective: To establish an ELISA detection method for serum antibody of mouse hepatitis virus (MHV) to conduct daily monitoring of MHV in this area in order to determine the infection status of SPF mice in time.

　　Method: Choose three MHV strains MHV1, MHV-JHM and MHV-A59, obtain purified and concentrated antigens through the proliferation and culture of L929 cells, and select the appropriate type of coating antigen. Immunize BALB/C mice to obtain high-titer immunopositive serum; optimize the ELISA reaction system to establish a standardized ELISA kit, and use it to detect clinical mouse serum samples.

　　Result: The MHV-coated antigen type suitable for this area was screened as MHV1, and a method for amplifying and concentrating the virus was established. The prepared MHV antiserum reaches multiple high titer levels in the same batch and can be used as a standardized quality control serum. The optimal concentrations of coated antigen, tested serum and enzyme complex are 4.0μg/mL (10-7.73/0.1 mLTCID50), 1:40 and 1:4000 dilutions, averaged within and between batches. The volatility is 5.13% and 5.57% respectively. The detection sensitivity is 1:4000 dilution. Mouse Sendai virus (SV), mouse pneumonia virus (PVM), type III Leovirus (Reo3), mouse parvovirus (MVM) and mouse poxvirus (Ect) positive sera have no cross-reactivity. The relative deviation of the stability test is less than 10%. 165 serum samples were tested, and the consistency rate of positive serum was 97.37% (37/38), and the consistency rate of negative serum was 92.19% (118/127).

　　Conclusion: The ELISA method established in this study has high specificity, sensitivity and reproducibility in the detection of MHV antibodies in mouse serum, and can be used for daily pathogenicity monitoring of mouse MHV in this area. You can accurately determine its infection status.