动物造模 z-bio 2022-04-23 306

　　Objective: To prepare mouse MHV, LCM, ECT and HANT liquid-phase chips, and to establish an xMAP multiplex detection method for transgenic mouse infectious disease antibodies.

　　Methods: The fluorescent microspheres were coupled with mouse MHV, LCM, ECT and HANT proteins respectively, and the optimal protein coupling amount, optimal working concentration of detection antibody and optimal working concentration of Streptavidin-PE were optimized. Mouse MHV, LCM, ECT and HANT liquid-phase chips were used for multiplex detection of xMAP on 56 transgenic mouse serum samples, negative serum, positive serum and quality control serum. And the traditional ELISA method was used to verify the detection results of xMAP.

　　Results: 1) The optimal coupling amounts of mouse MHV, LCM, ECT and HANT proteins were 20 μg, 20 μg, 40 μg and 20 μg, respectively, and the optimal concentrations of detection antibodies were 2 μg/mL and 2 μg/mL, respectively. , 2 μg/mL and 4 μg/mL; the optimal concentration of Streptavidin-PE antibody was 2 μg/mL; (2) xMAP test results showed that the serum MHV MFI of No. 14, No. 23, No. 55 and No. 56 transgenic mice The MFI value and index value of the serum of the other mice were lower than those of the quality control serum, indicating that the transgenic mice No. 14, No. 23, No. 55 and No. 56 were positive for MHV antibodies, and the rest of the mice were positive for MHV antibodies. MHV, LCM, ECT and HANT antibodies were negative; (3) ELISA test results showed that the MHV A450 value and index value of transgenic mice No. 14, No. 23, No. 55 and No. 56 were higher than those of the quality control serum. Both the A450 value and index value were lower than the quality control serum, indicating that the transgenic mice No. 14, No. 23, No. 55 and No. 56 were positive for MHV antibodies, and the other mice were negative for MHV, LCM, ECT and HANT antibodies. The ELISA test results were consistent with xMAP The test results are consistent.

　　Conclusion: The established xMAP multiplex detection technology can be applied to the detection of infectious diseases in transgenic mice.