猪戊型肝炎病毒吉林地区分离株全基因序列的分析

Whole gene sequence analysis of swine hepatitis E virus isolated in Jilin area

为了了解吉林地区HEV分子背景和流行特征,对2013年采自吉林地区养殖场的毒株编号为Ch-S-1的Ⅳ型戊型肝炎病毒(HEV)的全基因序列进行了分析。通过提取病毒RNA,采用RT-PCR方法分段扩增HEV全长基因,两端采用RACE法扩增,经过序列测定和拼接,利用ClusterX1.81、MEGA3.0、Phylop win 3.0软件分析基因序列。最终得到了HEV Ch-S-1全长基因组(GenBank登录号EF077630,7 261bp),其中ORF1、ORF2和ORF3分别编码1 706,674,114个氨基酸。与人源及猪源各基因型HEV相比,Ch-S-1与基因Ⅰ~Ⅲ型的ORF1片段同源性在78.8%~86.9%之间,而与基因Ⅳ型的ORF1片段同源性在93.7%~97.6%。Ch-S-1与基因Ⅰ~Ⅲ型的ORF2片段同源性在87.5%~92.4%之间,而与基因Ⅳ型的ORF2片段同源性在95.9~98.2%。Ch-S-1与基因Ⅰ~Ⅲ型的ORF3片段同源性在75.2%~84.0%之间,而与基因Ⅳ型的ORF3片段同源性在97.5%~99.2%。结果表明:HEV Ch-S-1全序列的基因结构特征与HEVⅣ型基本一致。

Hepatitis E is an important public health problem in several developing countries and causes the popularity of infection. This study was to understand the HEV molecular background and popular characteristics in Jilin area through the whole gene sequence analysis of HEV Ch-S-1. Viral RNA was extracted,and then HEV whole gene was amplified by RT-PCR and RACE. HEV whole gene was sequenced, spliced, and analyzed by ClusterX1. 81, MEGA3.0, Phylop win 3.0 software. The HEV Ch-S-1 full-length genomic （GenBank Accession No. EF077630,7 261 bp）, and ORF1,0RF2 and ORF3 encode 1706,674 and 114 amino acids were obtained. The amino acid homology of Ch-S-10RF1 and typeⅠ-Ⅲ ORF1 was 78.8%-86.9%,while 93.7%-97.6% with gype Ⅳ. The that of Ch-S-10RF2 and typeⅠ-Ⅲ ORF2 was 87.5%-92.4% ,while 95.9%-98.2% with typeⅣ. That of Ch-S-10RF3 and typeⅠ-Ⅲ ORF3 was 75.2%-84.0% ,while 97.5%-99.2% with type Ⅳ. HEV Ch-S-1 structure of the complete sequence of the gene is consistent with HEV Ⅳ type.