摘要
为建立快速检测塞内卡病毒A (Senecavirus A,SVA)血清学方法,以纯化的重组VP1蛋白作为包被抗原,建立了SVA VP1间接ELISA抗体检测方法。结果表明,VP1蛋白以包涵体形式表达,纯化后的蛋白经Western blot鉴定具有较好的反应原性。该ELISA检测方法阴、阳性临界值为0.278,与口蹄疫病毒(FMDV)、脑心肌炎病毒(EMCV)、猪圆环病毒2型(PCV2)、猪瘟病毒(CSFV)、猪伪狂犬病病毒(PRV)和猪繁殖与呼吸综合征病毒(PRRSV)等猪常见病原阳性血清均无交叉反应,具有良好的特异性。批内和批间重复性试验的变异系数均小于13%,表明该方法重复性和稳定性均较好。相比于血清中和试验(SN),该方法的相对敏感性为98.0%,两种方法的符合率为84%。用该方法对来自山东、广东省的8个猪场的276份血清进行检测,阳性率为22.1%。SVA VP1间接ELISA抗体检测法可作为一种快速、简便的血清学诊断方法,用于猪群SVA感染监测和流行病学初步调查。
In order to establish a rapid assay for detection of the serum antibodies against Senecavirus A(SVA),the indirect ELISA based on recombinant VP1 protein that expressed in BL21,E.coli was developed in this study.The result showed that protein VP1 had been expressed as the inclusion body.The purified protein has a good reactivity by Western blot.The critical value of this ELISA was 0.278.And this method had no cross reaction with other positive serum including FMDV,EMCV,PCV2,CSFV,PRV,PRRSV,which showed a good specification.The coefficient of variation(CV) of inter and intra-assay were both less than 13.0%,indicating a great repeatability and stability.The relative sensitivity was 98.0% compared with the SN test.The coincidence rate of the two methods was 84.0%.The positive rate of SVA antibodies was 22.1% in 276 clinical pig serums come from Shandong and Guangdong province.In a word,this indirect ELISA assay based on VP1 could be used for the monitor of infection as well as preliminary epidemiology survey,as a rapid and simple diagnosis method.
作者
范慧
曾洁
李亮
李仕海
张盼盼
延君芳
姜平
白娟
FAN Hui;ZENG Jie;LI Liang;LI Shi-hai;ZHANG Pan-pan;YAN Jun-fang;JIANG Ping;BAI Juan(Key Laboratory of Animal Diseases Diagnostic and Immunology of the Ministry of Agriculture ,Nanjing Agricultural University , Nanjing 210095 ,China)
出处
《中国兽医学报》
CAS
CSCD
北大核心
2019年第10期1907-1912,共6页
Chinese Journal of Veterinary Science
基金
国家自然科学基金资助项目(31502082)
中央高校基本科研业务费专项资助项目(Y0201800849,KJQN201616)