鼠肠道病毒RT-PCR检测与基因序列分析

RT-PCR detection of enterovirus carried by wild rodents and gene sequence analysis

目的对鼠类携带的肠道病毒进行检测并分析病毒基因序列特征。方法设计肠道病毒属一致-简并引物,采用RT-PCR方法对宁波口岸捕获的鼠类肠道内容物样品进行检测,对PCR产物进行测序,并将所测序列与国内外肠道病毒属病毒株序列进行核苷酸同源性分析和构建种系进化树。根据基因序列比对结果,采用鼻病毒通用引物进行验证。结果共检测165份鼠类样品,2份黄毛鼠样品经检测为阳性,扩增片段大小为378bp,标识为DXEV1601。片段的系统发生分析显示DXEV1601与人鼻病毒1B SC9813株位于同一进化分支,与23个不同肠道病毒属病毒流行株同源性相似度范围为50.3%～59.6%。经鼻病毒特异性引物扩增为阳性。结论宁波地区野生鼠类中存在鼻病毒感染。

To detect and phylogeneticaly analyze enterovirus carried by rodents,one pair of consensus-degenerate primers were designed to amplify the gene of enterovirus,and then RT-PCR was applied to detect enterovirus carried by rodents which captured from Ningbo port area.PCR products were sequenced.The homologous analysis and phylogenetic analysis among the sequence results and different enterovirus strains were performed.Using the Rhinovirus universal primers to verify the sample based on the sequence comparison.All 165 rodents’ samples were screened for enterovirus,2 Rattus loseatested positive,the amplified products was about 378 bp,marked as DXEV1601.The phylogenetic analysis showed that DXEV1601 were in one branch of phylogenetic tree with Human rhinovirus 1B strain SC9813.The nucleotide identity of DAEV1601 to other 23 enterovirus strains was from 50.3%to 59.6%.In conclusion,we prove the existence of Rhinovirus in wild rodents in Ningbo,China.