猪流行性腹泻病毒血清IgG间接ELISA检测方法的建立
投稿时间:2018-12-14    点此下载全文
引用本文:涂光岚,奉竹,桑益旸,李思思,王恒安.猪流行性腹泻病毒血清IgG间接ELISA检测方法的建立[J].《上海交通大学学报》(农业科学版),2019,(5):1-5, 9
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作者单位E-mail
涂光岚 上海交通大学 农业与生物学院,上海市兽医生物技术重点实验室,上海 200240 tuguanglan0123@sjtu.edu.cn 
奉竹 上海交通大学 农业与生物学院,上海市兽医生物技术重点实验室,上海 200240  
桑益旸 上海交通大学 农业与生物学院,上海市兽医生物技术重点实验室,上海 200240  
李思思 上海交通大学 农业与生物学院,上海市兽医生物技术重点实验室,上海 200240  
王恒安 上海交通大学 农业与生物学院,上海市兽医生物技术重点实验室,上海 200240  
基金项目:上海市科委攻关项目(15391901600)
中文摘要:以大肠杆菌表达系统制备的猪流行性腹泻病毒(porcine epidemic diarrhea virus,PEDV)中和抗原S1D(1 μg/mL)每孔100 μL进行包被,利用HRP-兔抗猪IgG(1∶20 000)建立了检测猪血清中抗PEDV IgG的间接ELISA方法。阴性判断临界值(M+2SD)为0.632,批内和批间检测的平均变异系数分别为2.3%和3.1%,该方法与商品化的PEDV抗体间接ELISA检测试剂盒同时检测50份临床血清样品,均为阳性,上述结果表明该方法可用于PEDV血清IgG的临床检测。
中文关键词:猪流行性腹泻病毒  IgG  中和抗原  间接ELISA
 
Establishment of an indirect ELISA for detecting serum IgG to porcine epidemic diarrhea virus
Abstract:E.coli-expressed porcine epidemic diarrhea virus(PEDV) neutralization antigen S1D(1 μg/mL,100 μL per well) and rabbit anti-pig IgG-HRP(1∶20 000) were used to establish an indirect ELISA for the detection of anti-PEDV IgG in pig serum.OD450=0.632 was set as the negative cut-off value(M+2SD),and the variation coefficients of intra-and inter batch were 2.3% and 3.1%,respectively.Fifty clinical sera samples were tested with the developed indirect ELISA using approved-commercial PEDV indirect ELISA kit as comparison and all samples were judged positive with each method.Taken together,these results indicated that the indirect ELISA in this study could be used for detecting antibodies to neutralizing antigen PEDV S1D.
keywords:PEDV  IgG  neutralizing antigen  indirect ELISA
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