猪传染性胃肠炎病毒HB-1株的分离鉴定和全基因组序列分析

Isolation,Identification and Whole Genome Sequence Analysis of Porcine Transmissible Gastroenteritis Virus HB-1 Strain

为了解河北省秦皇岛市某猪场猪传染性胃肠炎病毒(TGEV)的流行情况,本试验通过病毒分离培养、反转录PCR(RT-PCR)、间接免疫荧光试验(IFA)等方法进行病毒分离和鉴定,并分析其全基因组序列。结果显示,本试验从仔猪粪便样品中成功分离获得1株TGEV,命名为HB-1株。HB-1株全基因组核苷酸序列全长28 590 bp,含有9个开放阅读框,GenBank登录号为MZ368889;HB-1株与AYU和WH-1株全基因核苷酸序列相似性最高,同属Purdue谱系;与16个国内外毒株全基因核苷酸序列同源性介于94.5%~99.9%,与9个国内参考株全基因核苷酸序列同源性介于98.5%~99.9%。结果表明,本试验获得TGEV分离株与国内流行株亲缘关系较近,应为交叉感染导致,此结果为河北省TGEV分子流行病学调查提供参考数据。

In order to determine the prevalence of porcine transmissible gastroenteritis virus(TGEV)in a pig farm in Qinhuangdao city,Hebei province,this study undertook virus isolation and culture,reverse transcription PCR(RT-PCR)and indirect immunofluorescence assay(IFA)to isolate and identify the virus,and analyzed the whole genome sequence.The results showed that one TGEV strain was isolated from fecal samples of piglets and named as HB-1 strain.The total nucleotide length of HB-1 strain was 28590 bp,containing 9 open reading frames,and GenBank accession number was MZ368889.The nucleotide sequence of HB-1 strain shared the highest similarity with AYU and WH-1 strains of Purdue lineage.The nucleotide sequence homology was between 94.5%and 99.9%with 16 domestic and foreign strains,and between 98.5%and 99.9%with 9 domestic reference strains.Thus,TGEV HB-1 strain is closely related to the endemic strains in China,and may have originated from cross infection.These findings provide useful details on the molecular epidemiology of TGEV in Hebei province.