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?:abstract
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In order to evaluate the immune levels of type A FMD vaccine, the monoclonal antibody (MAb) 3D9 of FMDV prepared in our lab was used as the capture antibody, and the HRP- labeled MAb 9A9 was used as the detection antibody A solidphase competition ELISA method (SPCE) for the detection of antibody against FMDV serotype A based on monoclonal antibodies was developed and its conditions were optimized The results showed that the optimal concentration of MAb 3D9 was 1 16 g/mL, the optimal dilution of A- type FMDV antigen was 1:5, and the optimal dilution of HRP- labeled MAb 9A9 was 1:5000 When the serum diluted at 1:32, the critical value of the assay was determined to be 35% The method was used to detect positive serum of type A FMDV, type O FMDV, BCV, BRV, PRRSV, PCV and CSFV, respectively The results showed that the solid phase competition ELISA method could specifically detect type A FMDV, but had no cross- reaction with other viruses It showed that the method established in this study has strong specificity The sensitivity test was performed on 3 positive sera samples by VNT (virus neutralization test) The sensitivity is 1:1 024, 1:256, 1:128, respectively, indicating the method has high sensitivity Repeatability results showed the coefficient of variation of the intra-assay and inter-assay repeated tests were less than 10%, indicating that the repeatability is good This method was used to detect 112 serum samples along with the liquid- phase blocking ELISA method (LPBE) and VNT and analyze the correlation The results showed the correlation coefficient between the method established in this study and LPBE and VNT was 0 901 and 0 916, respectively It indicated that this method had a good reliability and has a higher correlation with VNT Using the SPCE and Korean Jeno A FMDV ELISA antibody detection diagnosis kitto detect 470 clinical samples (90 sheep serum samples, 170 cow serum samples and 210 pig serum samples) The results showed the overall coincidence rate between this method and Jeno kit is 90 3%, 91 8% and 89 0%, respectively, indicating that the detection result of this method is relatively accurate This study laid a foundation for the establishment of a method for the detection of immunity level of type A foot-and-mouth disease vaccine
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