烟台沿海地区人源和猪源戊型肝炎病毒基因型别的研究

摘要

目的 调查烟台市沿海地区人源与猪源戊型肝炎病毒(HEV)基因型别的相关性.方法 应用逆转录一巢式聚合酶联反应( RT-nPCR)方法对当地急性散发戊型肝炎患者、正常人群中抗HEV-IgM阳性者和当地养猪场的猪进行HEV RNA检测,并对HEV RNA阳性标本进行克隆测序和序列分析.结果 16例急性散发戊型肝炎患者中有7例粪便标本HEV RNA阳性;51份lgM阳性正常人群血清标本中有1份HEV RNA阳性;34份猪胆汁标本中有1份HEV RNA阳性.序列分析发现该地区HEV人株与猪株在ORF2部分区域的核苷酸序列同源性为87％～ 98.1％.7株患者的戊肝病毒基因型和1株猪的戊肝病毒基因型均为Ⅳ型,基因序列同源性在87％ ～ 98.1％之间;其中有6例患者和猪的基因序列同源性在93.9％ ～98.1％之间,为Ⅳ型a亚型;1例患者和猪的基因序列同源性为87％,为Ⅳ型d亚型.正常人群的1例戊肝病毒基因型为Ⅰ型d亚型.该地区人与猪HEV的ORF2的部分基因片段与HEV Ⅰ～Ⅳ型的代表株进行比较,核苷酸序列同源性分别是82.5％～100％,81.7％ ～92.9％,81.4％ ～93.9％,84.9％ ～ 100％.结论 该地区人群中流行的HEV存在2个基因型3个亚型,主要以基因Ⅳa型为主,与猪群中流行的HEV基因Ⅳa型同源性较高;HEV Ⅰ型在人群中散在存在.%Objective Survey of the coastal city of Yantai,from human and swine hepatitis E virus (HEV) genotype correlation. Method Application of reverse transcription nested polymerase chain reaction (RT - nPCR) method for local acute sporadic hepatitis E patients,normal population of HEV-IgM positive and local pig farm pigs were HEV RNA detection.And HEV RNA positive samples for cloning sequencing and sequence analysis.Results In 16 patients with acute sporadic hepatitis E in 7 cases of RNA positive stool specimens of HEV; 51 lgM positive sera of normal people in specimens with 1 HEV RNA positive; 34 pig bile specimens with 1 HEV RNA positive. Sequence analysis revealed the region HEV strains and swine strains in the ORF2 region of nucleotide sequence homology is 87％ -98.1％.7 strains of hepatitis E virus genotype in patients and 1 strains of swine hepatitis E virus genotypes are type Ⅳ,gene sequence homology between the 87％ -98.1％ ; there were 6 patients and porcine gene sequence homology in 93.9％ -98.1％ between,for type a subtype; 1 patients and porcine gene sequence homology in 87％,for the type D subtype.Normal population of 1 cases of hepatitis E virus genotype for I type D subtype.Human and porcine HEV ORF2 gene fragment and HEV part Ⅰ-Ⅳ representative strains were compared,and the nucleotide sequence homology were 82.5％ - 100％,81.7％ - 92.9％,81.4％ - 93.9％,84.9％ - 100％.Conclusion The area population prevalence of HEV in the presence of 2 genotype 3 subtype genes,mainly to Ⅳ A,in pigs with popular HEV gene with a high homology; HEV type Ⅰ in the crowd disperses in the presence of.