猪流行性腹泻病毒重组N蛋白间接ELISA检测方法的建立  

An indirect ELISA for the detection of antibodies against porcine epidemic diarrhea virus using recombinant N protein as coating antigen

作　　者：姜艳平[1] 姜新鹏[1] 孙刘妹[1] 赵广宇[2] 崔文[1] 唐丽杰[1] 乔薪瑗[1] 李一经[1] 

机构地区：[1]东北农业大学动物医学学院,黑龙江哈尔滨150030 [2]东北农业大学园艺学院,黑龙江哈尔滨150030

出　　处：《中国预防兽医学报》2015年第2期119-122,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金："十二五"农村领域国家科技计划课题(2012AA101304-3)

摘　　要：为建立检测猪流行性腹泻病毒(PEDV)血清抗体的间接ELISA方法,本研究以原核表达的重组N蛋白作为检测抗原,优化ELISA反应条件,建立了PEDV间接ELISA抗体检测方法。该方法仅对PEDV血清检测为阳性,对猪传染性胃肠炎、猪轮状病毒感染猪的阳性血清检测结果均为阴性;其批内和批间重复性试验的变异系数均小于10%。采用建立的ELISA方法检测了130份临床样品,其中78份血清样品为PEDV抗体阳性。表明建立的以重组N蛋白为包被抗原检测PEDV血清抗体的方法可以用于检测PEDV感染及相关的流行病学调查。Porcine epidemic diarrhea virus （PEDV） is one of the most important pathogens causing severe enteritis and watery diarrhea in piglets. To establish the serum diagnosis method of PEDV infection, an indirect ELISA was developed for the detection of PEDV antibody with the N protein of PEDV as coating antigen expressed in E.coli. Under the optimized reaction conditions, the assay was able to specifically detect the positive serum of PEDV, but no cross-reactions were found for the positive sera of other virus, including porcine transmissible gastroenteritis virus and porcine rotavirus. The coefficients of variation for intra and inter-assay were lower than 10%. A total of 130 clinical serum samples tested, of which 78 were positive for antibodies against PEDV. These results indicate that the indirect ELISA is a rapid, sensitive and specific for detecting antibodies against PEDV and has a potential to apply for diagnosis of PED and for serologic survey of PEDV infection.

关 键 词：猪流行性腹泻病毒 重组N蛋白 抗体检测 诊断方法 

分 类 号：S852.65[农业科学—基础兽医学]