中文摘要：

为建立快速检测脑心肌炎病毒（EMCV）的环介导等温扩增方法（LAMP）,参照GenBank中EMCV基因组序列（X74312.1）,针对EMCV的3D基因保守区域设计并合成了4条引物,对建立的反应体系进行条件优化,并进行特异性、敏感性试验及临床样本的检测。结果显示,成功建立了EMCV LAMP检测方法,且当内、外引物浓度比例为5∶1（即内、外引物浓度分别为50μmol/L、10μmol/L）,反应温度为64℃,反应时间为60min时反应体系可达最佳。特异性和敏感性试验表明,该方法能够特异性地检测EMCV,且敏感性比常规RT-PCR法高10倍。分别用建立的LAMP法与ELISA法对临床样本进行检测,结果二者的符合率为97%。表明建立的LAMP方法特异性强、敏感性高,可适用于EMCV临床样本的快速检测。

英文摘要：

To establish a loop-mediated isothermal amplification （LAMP） technology, four primers were designed and synthesized,which was focused on the conserved region of 3D gene,retrieved from GenBank（X74312.1）. Using these four primers to optimize the reaction system conditions,its specificity, sensitivity and clinical samples were assessed. In result, the LAMP technology was successfully established. LAMP results showed to be optimal when the inner and outer primer concentration ratio was 5 ： 1（inner and outer primers concentrations were of 50μmol/L, 10μmol/L respectively）, the reaction temperature was 64 ℃ and reaction time was 60 min. This technology was specific for EMCV and its sensitivity was ten times higher than conventional RT-PCR. By detecting 100 clinical samples by LAMP and ELISA,respectively,the coincidence of both methods was 97%. It suggests that the LAMP method is with strong specificity and high sensitivity and can be applied to the rapid detection of EMCV clinical specimen.