摘要
为了解河北地区牛群牛病毒性腹泻病毒(bovine viral diarrhea virus,BVDV)抗体阳性率,本研究利用E.coli原核表达系统表达了E^(rns)蛋白。以该蛋白为包被抗原,通过优化各反应条件,建立了检测BVDV抗体的间接ELISA方法,并利用该方法检测河北地区477份牛血清样品的BVDV抗体阳性率。特异性试验结果显示,本研究建立的BVDV抗体ELISA检测方法仅能检出BVDV抗体阳性血清,而检测牛传染性鼻气管炎病毒阳性血清、牛布鲁菌阳性血清、牛轮状病毒阳性血清、牛冠状病毒阳性血清、口蹄疫病毒阳性血清均为阴性。敏感性试验结果显示,建立的ELISA方法检测BVDV阳性血清具有较高的敏感性。重复性试验结果显示,ELISA检测方法的批内和批间变异系数均小于10%。应用本研究建立的抗体ELISA检测方法与IDEXX试剂盒检测某牛场90份血清样品,总符合率为93.33%。对河北地区18个牛场的477份血清样品进行检测,结果接种疫苗牛场的血清阳性率为30.43%~92.86%,未接种免疫牛场抗体阳性率为0%~100%。结果表明,本研究建立的BVDV抗体ELISA检测方法具有较高的特异性、敏感性和重复性,可应用于临床牛血清样品检测,为河北地区BVDV的监测和防控提供技术支持。
To reduce the cost of BVDV antibody detection and determine the BVDV antibody positive rate of cattle in Hebei Province,the E.coli prokaryotic expression system was used to express the protein of E^(rns).An indirect ELISA method for detection of BVDV antibody was established by optimizing the reaction conditions.The positive rate of BVDV antibody in 477 bovine serum samples from Hebei Province was detected by this method.The specificity test results showed that the BVDV antibody ELISA established was positive for BVDV antibody-positive serum,but negative for bovine infectious rhinotracheitis virus positive serum,bovine Brucella positive serum,bovine rotavirus positive serum,bovine coronavirus positive serum,foot and mouth disease virus positive serum.The results of the sensitivity test showed that the ELISA established was highly sensitive to BVDV positive serum(the maximum dilution of positive serum was 1600 times).The results of the repeatability test showed that the coefficient of variation of the ELISA method was less than 10%.The total coincidence rate between the ELISA method and the IDEXX kit was 93.33%.477 serum samples from 18 cattle farms in Hebei Province were detected by this method.The positive rate of vaccinated cattle farms was 30.43%-92.86%,and that of non-vaccinated cattle farms was0%-100%.The results showed that the ELISA method established in this study had high specificity,sensitivity and repeatability,and could be applied to the detection of clinical bovine serum samples,providing technical support for the monitoring and prevention of BVDV in Hebei Province.
作者
刘勃兴
刘畅
王利丽
付祥
吴同垒
刘洁
高荣菊
张志强
史秋梅
LIU Boxing;LIU Chang;WANG Lili;FU Xiang;WU Tonglei;LIU Jie;GAO Rongju;ZHANG Zhiqiang;SHI Qiumei(Hebei Key Laboratory of Preventive Veterinary Medicine,Hebei Normal University of Science&Technology,Qinhuangdao,Hebei 066004,China;Agricultural and Rural Bureau of Leting County,Tangshan,Hebei 063600,China)
出处
《中国兽医学报》
CAS
CSCD
北大核心
2022年第3期445-451,471,共8页
Chinese Journal of Veterinary Science
基金
河北省农业厅资助项目(HBCT2018130203)
河北省重点研发计划资助项目(19226629D)
河北省委组织部资助项目(180416)。
