动物医学进展2024，Vol.45Issue(1)：44-51,8.
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
Establishment and Application of Indirect Immunofluorescence Assay for Detecting Egg-drop Syndrome Virus
摘要
Abstract
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.关键词
鸡产蛋下降综合征病毒/多抗血清/间接免疫荧光/优化Key words
EDSV/polyclonal antiserum/indirect IFA/optimization分类
农业科技引用本文复制引用
孔冬妮,薛青红,李俊平,毛娅卿,王嘉,邓永,翟天舒,刘丹,黄小洁,薛琪,候力丹,吴华伟..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[J].动物医学进展,2024,45(1):44-51,8.基金项目
中国兽医药品监察所公益项目(GY202105) （GY202105）
