Abstract: Abstract: Objective To develop a real-time ＲT-PCＲ assay ( rＲT-PCＲ) for efficient detection of Murine Encephalomyocarditis virus ( EMV) ． Method According to the genomic sequences of EMV strain PEC9 ( DQ288856. 1) download from NCBI and to find the conserved sequences． One pair of the specific primers and one TaqMan probe were designed． Then reaction parameters were optimized to develop a real-time ＲT-PCＲ assay ( rＲTPCＲ) ． Ｒesult The method showed a good linear relationship of standard curve with a r 2 value of 0. 99． The sensitivity of the real-time PCＲ was less than 1 copies /μL． 1 000 times higher than ordinary PCＲ method ，and its specificity is strong，common mice strains had no specific amplification，between batch and batch of variation coefficient is less than 2% ． 120 mice feces samples epidemic materials provided by the Shanghai institute of experimental animals were detected by the real-time PCＲ． All the 120 mice samples were negative． Conclusion The real-time ＲT-PCＲmethod is good in specificity and sensitivity，which can make a powerful technical support for EMV epidemiological investigation and detection．

Key words: <p>Murine Encephalomyocarditis virus, EMV, TaqMan, real time PCＲ</p>

摘要： 摘要: 目的 建立快速诊断鼠脑心肌炎病毒感染的荧光定量 ＲT-PCＲ 方法。方法 根据鼠脑心肌炎病毒( EMV) PEC9 毒株全基因组序列( GenBank 登录号: DQ288856. 1) ，设计一对特异性引物和 TaqMan 探针，建立 EMV 的 TaqMan 探针荧光定量 ＲT-PCＲ 方法，进行条件优化，检测其特异性，敏感性，稳定性。结果 建立的荧光定量 PCＲ方法，标准曲线的线性关系良好，r 2 值可达 0. 99，敏感性最低检测限为 1 个拷贝数，高于普通 PCＲ 方法 1000 倍，其特异性强，与其他常见感染的小鼠病毒株均无特异性扩增，批内和批间变异系数均小于 2% 。利用该方法对上海市 120 份小鼠心肌样品进行检测，未检测到阳性感染。结论 本研究为鼠脑心肌炎病毒感染的分子检测，为制定国家标准提供良好的方法。

关键词: <, p>, 鼠脑心肌炎病毒, EMV, TaqMan 探针, 荧光定量 ＲT-PCＲ<, /p>