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目的测定从新疆南部地区分离的猪戊型肝炎病毒(HEV)株全基因组序列,并在此基础上分析猪HEV与人源HEV的关系。方法设计HEV基因4型通用PCR引物,用逆转录巢式聚合酶链反应法(RT-nPCR)分段扩增猪HEV株CHN-XJ-SW33的全基因组序列;用cDNA末端快速扩增法(RACE)扩增其末端序列;对扩增的目的片段进行克隆测序,并对拼接后的基因组进行序列比对和进化分析。结果除3′poly(A)尾外,CHN-XJ-SW33基因组全长为7 238 nt,由3个开放读码框(ORF1-3)组成,分别编码1 706、674和114个氨基酸。CHN-XJ-SW33全基因组序列与HEV基因1-3型病毒株同源性仅为72.1%~74.9%,而与HEV基因4型病毒株同源性高达82.8%~95.5%,其中与日本人源中国输入型HEV株JKO-ChiSai98C同源性最高,为95.5%。基因进化分析显示,CHN-XJ-SW33属于HEV4a基因亚型。结论猪HEV与人HEV在全基因组核苷酸序列上高度同源,提示基因4型HEV可能由猪传染给人。
Objective To determine the whole genome sequence of swine hepatitis E virus (HEV) isolated from southern Xinjiang and to analyze the relationship between swine HEV and human HEV. Methods The universal HEV genotype 4 PCR primers were designed and the whole genome sequence of CHN-XJ-SW33 was amplified by RT-nPCR. The rapid amplification of cDNA ends RACE). The amplified fragments were cloned and sequenced, and the genomes were sequenced and analyzed. Results Except for the 3’poly (A) tail, the full length of CHN-XJ-SW33 gene was 7 238 nt in length and consisted of 3 open reading frames (ORF1-3) encoding 1 706, 674 and 114 amino acids, respectively. The genome sequence of CHN-XJ-SW33 shared only 72.1% -74.9% homology with HEV genotype 1-3 strains, while its homology with HEV genotype 4 strains was as high as 82.8% -95.5% The highest homology of JKO-ChiSai98C from China imported HEV strain was 95.5%. Gene evolution analysis showed that CHN-XJ-SW33 belongs to HEV4a gene subtype. Conclusion HEV and human HEV are highly homologous to the nucleotide sequence of the whole genome, suggesting that HEV may be transmitted from pigs to humans.