O型、A型及Asia1型3个型口蹄疫病毒通用套式RT-PCR检测方法的建立  

Development of a nest RT-PCR for simultaneously detecting serotype O, A and Asia1 FMDV

作　　者：李金海[1,2] 李兴玉[3] 曹三杰[2] 陈斌[1] 文心田 张毅[1] 

机构地区：[1]四川省动物疫病预防控制中心,四川成都610041 [2]四川农业大学动物医学学院/猪病研究中心,四川雅安625014 [3]四川省畜牧科学研究院,四川成都610066 [4]不详

出　　处：《中国预防兽医学报》2014年第10期784-787,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：公益性行业(农业)科研专项(201203056)

摘　　要：为了建立针对流行于我国及周边国家主要口蹄疫病毒(FMDV)血清型的检测方法,本实验设计2对特异性引物,通过对反应条件的优化,建立了能够同时扩增O型、A型及Asia1型FMDV VP1全基因的套式RT-PCR(RT-nPCR)方法。该方法能特异性检测O型、A型和Asia1型FMDV,对其它相关动物病毒的检测结果均为阴性;比FMDV多重RT-PCR商品试剂盒敏感约1 000倍。利用该方法对10份FMDV样品进行扩增检测,结合扩增产物的克隆测序技术,对所检样品进行了准确定型。实验结果表明,该方法通用型强,可用于3个血清型FMDV的检测和分子流行病学调查。In order to establish the assay for detecting the main serotypes of foot-and-mouth disease virus （FMDV） prevalent in China and neighboring countries and regions, a nest RT-PCR assay （RT-nPCR） was developed for detections of three serotypes of FMDV with two pair of primers targeting the VP1 gene. The results showed that the RT-nPCR was able to specifically amplify the fragwets from VP1 gene of A, O and Asial serotypes of FMDV, whereas, no amplified product was found from other related viruses. Moreover, the sensibility of established RT-nPCR was 1,000-fold higher than that of commercial mRT-PCR kit which was widely used in China. These data demonstrated that the RT-nPCR had the potential application for the diagnosis of FMDV and epidemiological investigations.

关 键 词：口蹄疫病毒 RT-nPCR VP1基因 

分 类 号：S852.65[农业科学—基础兽医学]