作 者：
刘邓;贾泽颖;张丹;全炎铭;郭又春;刘莉如;刘修全;陈华林
关键词：
传染性胃肠炎病毒(TGEV);猪流行性腹泻病毒(PEDV);轮状病毒(RV);检测;多重实时荧光定量PCR
摘 要：
为了建立一种能够同时检测猪流行性腹泻病毒(PEDV)、传染性胃肠炎病毒(TGEV)和轮状病毒(RV)的多重实时荧光定量PCR方法,试验根据PEDV CV777株、TGEV Purdue P115株全基因序列和RV OSU毒株的VP6基因序列,设计出3对特异性引物及探针,优化反应条件,开展特异性、敏感性、重复性和验证试验。结果表明:该方法的检测最低限为1×10~2拷贝/μL的RNA标准品或1×10~(-2)TCID_(50)/mL的疫苗毒,具有较高的敏感性;变异系数均小于10%,符合统计学要求;利用本试验所建立的方法以及商品化试剂盒对20份临床样品进行检测,符合率为100%。说明本试验建立的方法特异性好,敏感性高,重复性稳定,可用于PEDV、TGEV和RV的鉴别诊断和防控。
译 名：
Establishment of a multiplex real-time fluorescent quantitative PCR assay for simultaneous detection of porcine epidemic diarrhea virus,transmissible gastroenteritis virus and rotavirus
作 者：
LIU Deng;JIA Zeying;ZHANG Dan;QUAN Yanming;GUO Youchun;LIU Liru;LIU Xiuquan;CHEN Hualin;Animal Husbandry&Veterinary Station of Beijing San Yuan Group;
关键词：
Transmissible gastroenteritis virus(TGEV);;Porcine epidemic diarrhea virus(PEDV);;Rotavirus(RV);;test;;multiplex real-time fluorescence quantitative PCR
摘 要：
Three pairs of primers and props were designed respectively based on the sequences of PEDV CV777,TGEV Purdue P115 and RV OSU strain VP6 gene. A sensitive,specific,reproduc-eble and multi real-time fluorescence quantitative PCR method for simultaneous detection of PEDV,TGEV and RV was establishedthrough theoptimization of reaction conditions. The detection limit of the method was 1 ×10~2 copies/μL RNA standard or 1 × 10~(-2)TCID_(50)/mL vaccine poison,with high sensitivity. The coefficient of variation was less than 10% in the repeatability test,which met the statistical requirements; 20 clinical samples were detected by themethod established in thisstudy and commercial kit,the coincidence rate was 100% . This method displayed the advantages of good specificity,high sensitivity,repeatability and stability. It can be used for differential diagnosis,prevention and control of PEDV,TGEV and RV.
