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为探索东北地区戊型肝炎病毒(HEV)流行背景和特征,本实验用抗-HEV抗体酶联免疫试剂盒检测了不同样品血清中的抗体;对样品用RT-PCR检测HEV RNA,对PCR阳性产物进行克隆测序,然后对序列进行分析。对49个呈RNA阳性的样品进行了序列测定,结果显示HEV分离株之间核苷酸同源性为82.2%~100.0%;同时,还与其它四种基因型戊型肝炎病毒进行了核苷酸序列同源性比对,分别在75.6%~81.6%、77.7%~80.9%、73.6%~81.0%和80.9%~95.7%之间。本实验还针对吉林省内的动物源HEV与人源HEV的序列进行了比对,结果其同源性在86.2%~100.0%之间,其中有5对序列完全相同。本研究获得的44个核酸片段结合HEV基因Ⅳ型中已知亚型的12个基因序列进行了亚型分类。结果 44个核酸片段对应3个分支,包括Ⅳa、Ⅳb亚型及另一支独立分支,表明本实验所获得的独立分支里包含的病毒分离株可形成新的亚型,本实验中将其命名为Ⅳh亚型。中国东北地区主要流行的HEV基因型为Ⅳ型,并且人源HEV与动物源HEV有极高的同源性,同时,还发现中国东北地区流行着国内已有报道的Ⅳa、Ⅳb亚型外,存在着一个新的亚型。
In order to explore the epidemiological background and characteristics of hepatitis E virus (HEV) in Northeast China, anti-HEV antibody ELISA kit was used to detect the antibody in different samples. HEV RNA was detected by RT-PCR, The product was cloned and sequenced, and the sequence analyzed. Sequence analysis of 49 RNA-positive samples showed that nucleotide homology between HEV isolates ranged from 82.2% to 100.0%. At the same time, nucleosides of other four genotypes of Hepatitis E virus The homologies were between 75.6% -81.6%, 77.7% -80.9%, 73.6% -81.0% and 80.9% -95.7%, respectively. In this experiment, the sequences of HEV and human HEV in animal origin in Jilin province were also compared, and the homology was between 86.2% and 100.0%, of which 5 pairs were identical. The 44 nucleic acid fragments obtained in this study were sub-classified according to 12 gene sequences of known subtypes of type IV of HEV gene. Results The 44 nucleic acid fragments corresponded to three branches, including subgroups IVa and IVb and another independent branch, indicating that the virus isolates contained in the independent branches obtained in this experiment could form new subtypes. In this experiment, they were named Ⅳ h subtype. The prevalence of HEV genotype in northeastern China is type Ⅳ, and human HEV has very high homology with animal HEV. At the same time, it is also found that there are epidemiological reports of subtypes Ⅳa and Ⅳb that have been reported in northeastern China, There is a new subtype.