动物实验,肝炎病毒,ELISA z-bo 2020-10-12 275

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

　　Method: Select three types of 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. Immune 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: We screened the MHV-coated antigen type suitable for this region as MHV1 and established a virus amplification/concentration purification method. The prepared MHV antiserum reaches multiple high titer levels in the same batch and can be used as a standardized quality control serum. In coating, between batches and between batches, the best working concentration of the test serum and enzyme conjugate is 4.0μg/mL (10-7.73/0.1 mLTCID50), 1:40 and 1:4000 dilution . The average volatility coefficient is different. 5.13% and 5.57%, detection sensitivity 1:4000 dilution, mouse Sendai virus (SV), mouse pneumonia virus (PVM), type III Leovirus (Reo3), mouse parvovirus (MVM), 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, the serum consistency rate was 97.37% (37/38), and the negative serum consistency rate was 92.19% (118/127).

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