摘要
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 and phylogenetically analyze the complete genome of the swine hepatitis E virus（HEV）isolate CHN-XJ-SW33 isolated from southern Xinjiang,China.Methods The overlapping fragments of the HEV isolate CHN-XJ-SW33 were amplified with a reverse-transcription-nested polymerase chain reaction（RT-nPCR） and 5′ and 3′ ends of viral genome were amplified with a rapid amplification of cDNA ends（RACE）.The PCR products were cloned and sequenced.The sequence and phylogenetic analysis of the swine HEV isolate was performed.Results The full genome of the CHN-XJ-SW33 strains consisted of 7 238 nucleotides,excluding the poly（A） tail,and contained three ORFs（ORFs 1-3） that encoded proteins of 1 706,674 and 114 aa,respectively.The full genomic sequencing showed that the CHN-XJ-SW33 strains shared similarities with all known HEV genotype 1,2 and 3 isolates by 72.1%-74.9% and with an identity of 82.8%-95.5% among genotype 4 HEV isolates.CHN-XJ-SW33 shared a high nucleotide identity of 95.5% with human HEV strain JKO-ChiSai98C and located in the branch of HEV subtype 4a.Conclusion The high nucleotide similarity between human and swine HEV strains based on full genome suggests the possibility of swine to human transmission of genotype 4 hepatitis E virus.
出处
《中国预防医学杂志》
CAS
2010年第5期433-438,共6页
Chinese Preventive Medicine
基金
国家自然科学基金资助项目(30570063)
国家863高科技研究发展项目(2006AA02Z453)
关键词
戊型肝炎病毒
猪
基因型
全基因组序列
Hepatitis E virus（HEV）
Swine
Genotype
Full-length genome
