猪流行性腹泻病毒FJ-11A株的分离与ORF3基因序列分析

摘要

Porcine epidemic diarrhea virus (PEDV) FJ-11A strain was isolated from piglets suffering from a severe diarrhea in a pig farm of Fujian Province using Vero cell. The PEDV ORF3 gene was obtained using reverse transcription-polymerase chain reaction (RT-PCR) method according to the sequence download from the GenBank. The results showed that the ORF3 gene of the FJ-11A strain was 675 bp, encoding an open reading frame (ORF) with 224 amino acids. Compared with the ORF3 gene published previously in GenBank by DNAstar software, the ORF3 sequence of FJ-11A strain had the highest nucleotide homology at 99.3% with CPF1074 and PFF513, isolating in South Korea, and also had 97. 8% homology with vaccine strain CV777. Based on genetic evolutionary, the PEDV strains which isolated in China were in the same phylogenetic relationship, whereas the candidate vaccine strains (attenuated DR13 and CV777) were in the other phylogenic lineage.%从福建省某疑似流行性腹泻病毒感染的猪场采集腹泻粪便样品和小肠组织,利用Vero细胞连续传代后,分离到猪流行性腹泻病毒福建株(FJ-11A株).根据GenBank中登录的猪流行性腹泻病毒ORF3基因特征设计特异性引物,采用RT-PCR方法对猪流行性腹泻病毒福建株ORF3基因进行扩增,将扩增目的片段经胶回收后进行克隆测序.结果表明,所扩增的目的片段基因编码有完整的ORF3开放阅读框,全长为675 bp,编码有224个氨基酸.运用DNAStar软件,将获得的ORF3基因序列和GenBan中登录的猪流行性腹泻病毒株ORF3基因序列进行分析比较和遗传进化分析,猪流行性腹泻病毒福建株ORF3基因和韩国分离株CPF1074和PFF513该基因核苷酸同源性最高,均为99.3％,与疫苗株CV777核苷酸同源性为97.8％.从遗传进化上看,我国PEDV分离株ORF3基因处在同一个分支,而弱毒株则处在相对独立的分支.