鸡产蛋下降综合征病毒间接免疫荧光检测方法的建立及应用OACSTPCD

To establish an indirect immunofluorescence assay for the detection of Egg drop syndrome virus(EDSV)in chicken,the various steps of the assay were optimized.In this study,the combination of polyan-tiserum of chicken egg drop syndrome and FITC-labeled rabbit anti-chicken IgG was used to optimize the optimal working concentration and incubation time of the primary and secondary antibodies,and the sensi-tivity,specificity and repeatability of indirect IFA method were tested.The results showed that the optimal working concentration of polyclonal antiserum in 96-well cell culture plates inoculated with EDSV was 1∶320,the incubation time was 45 min.The optimal working concentration of FITC-labeled rabbit anti-chick-en IgG was 1∶200,incubation time was 30 min.Virus sensitivity test showed that the method could detect 10-7.0 TCID50/0.1 mL.The polyclonal antiserum sensitivity test showed that the specific fluorescence was still obvious when the polyclonal antiserum was diluted to 1∶1 280.Moreover,the generated polyclonal an-tiserum against EDSV only targeted at EDSV,did not show cross-reaction with 8 other avian pathogens(FAdV 4,NDV,IBDV,IBV,REV,ALV,MDV,AIV).The results of intra-batch and inter-batch repeat-ability tests showed that the method has good repeatability and reliability.Overall,the preliminary estab-lishment of indirect immunofluorescence assay in this stusy has high sensitivity and stability,also provides technical support for the detection of poultry biological material EDSV and has an important market de-mand.