南方农业学报2011，Vol.42Issue(3)：324-327,4.
猪脑心肌炎病毒一步法RT-PCR检测方法的建立及应用
Development of one-step RT-PCR assay for detection of porcine ence-phalomyocarditis virus and its applications
摘要
Abstract
[Ohjective ]The present experiment was aimed to develop a rapid and accurate method for the detection of porcine encephalomyocarditis (EMCV) pathogen and its clinical diagnosis. [Method]A one-step reverse transcription-polymerase chain reaction (RT-PCR) for detection of porcine encephalomyocarditis virus (EMCV) was developed using a pair of specific primers based on the 3D gene sequences of EMCV strains obtained from GenBank. The specificity, sensitivity and repetition of the developed method were also conducted. [Result]The developed one-step RT-PCR could specifically amplify a 286 bp fragment f'rom EMCV BJC3 strain. The test gave negative results for the strains of foot-and-mouse disease virus (FMDV) , classical swine fever virus (CSFV) , porcine reproductive and respiratory syndrome virus (PRRSV) , porcine pseudorabies virus(PRV) , porcine parvovirus (PPV) and normal BHK-21 cells. The detection limit of the developed assay was determined to be as little as 2 tissue culture infectious dose (TCID50) of EMCV. Repeated detection from 10 EMCV-infected cell cultures by the one-step RT-PCR resulted in the same results. The assay was used for 207 clinical samples from suspected piglets, of' which 19 gave the positive results, with 9.18％ of positive rate. [Conclusion] The established onestep RT-PCR assay was specific , sensitive , eff'icient and rapid for the clinical detection and epidemiological investigationof EMCV infection.关键词
猪/脑心肌炎病毒(EMCV)/3D基因/一步法RT-PCRKey words
porcine / encephalomyocarditis virus/ 3D gene / one-step RT-PCR分类
农业科技引用本文复制引用
施开创,莫胜兰,胡丽萍,杨荣,郑敏,李军..猪脑心肌炎病毒一步法RT-PCR检测方法的建立及应用[J].南方农业学报,2011,42(3):324-327,4.基金项目
广西青年科学基金项目(桂科青0728047) （桂科青0728047）
广西科技创新能力与条件建设基金项目(08-05-01D) （08-05-01D）
