猪流行性腹泻病毒的RT-PCR鉴定及其 M、N和E基因的序列分析

对引起仔猪严重腹泻的猪流行性腹泻病毒(Porcine epidemic diarrhea virus,PEDV)进行RT-PCR检测,并对PEDV部分基因进行克隆和序列分析。从陕西省某发生严重仔猪腹泻的养猪场采集肠道内容物,利用RT-PCR技术成功检测到PEDV感染。克隆检测到的PEDV M、N和E基因片段,并将该基因核苷酸序列和推导的氨基酸序列与其他不同来源的PEDV毒株进行比较分析。结果表明,克隆的M、N和E基因与其他已经发表的PEDV毒株核苷酸的同源性分别为96.3%～99.9%、95.2%～99.5%和96.1%～96.9%,氨基酸同源性分别为95.6%～99.6%、96.1%～99.5%和96.1%～98.7%;系统进化树结果表明,试验检测的PEDV与国内分离株的PEDV株亲缘关系更近。可见:陕西省养殖场存在猪流行性腹泻病毒感染,其M、N和E基因变异不大。

RT-PCR Identification of Porcine Epidemic Diarrhea Virus and Sequence Analysis of Its M, N and E Genes

To identify a porcine epidemic diarrhea virus (PEDV) strain which causes a severe piglet with diarrhea by RT-PCR method, and part genes of PEDV was cloned and analyzed its phylogenetic relationship with other PEDV strains. Intestinal tract contents of a severe piglet with diarrhea were collected from a pig farm in Shaanxi province and a porcine epidemic diarrhea virus (PEDV) strain was identified by RT-PCR method. The M, N and E genes of PEDV identified were cloned and compared nucleotide sequence and the homology of deduced amino acids sequence with other PEDV strains. Compared with other published PEDV strains, the homology of M, N and E genes of PEDV identified were 96.3%-99.9%, 95.2%-99.5% and 96.1%-96.9%, respectively. The homology of deduced amino acids sequence between PEDV identified and other strains were 95.6%-99.6%, 96.1%-99.5% and 96.1%-98.7%, respectively. Phylogenetic tree analysis showed that the identified strain was genetically close to PEDV strains from China. There exists porcine epidemic diarrhea viruses in Shaanxi province, and the differences of M, N and E genes between the identified strain and other strains are not obvious.