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4例禽戊型肝炎病例的实验室诊断及ORF2基因遗传进化分析

点击数:7806  时间:2019-08-13 16:51:16  来源: 信息处      

本研究通过分子流行病学诊断技术,分别从山东、辽宁、吉林、陕西4个地区表现肝炎症状的病鸡中,采集肝脏和脾脏样品,利用RT-PCR方法,检测禽戊型肝炎病毒(aHEV),并对aHEV-ORF2 基因片段进行测序,利用MegAlign进行同源性比较。结果显示:4个鸡场的病鸡样品均为aHEV 阳性,aHEV-ORF2基因序列间同源性为84.5%~94.5%,与已报道的4种基因型同源性均小于84.0%,其中与基因1型的澳大利亚株和韩国株同源性为81.6%~82.8%,与基因2型的美国原型株和美国无毒株同源性为80.9%~82.8%,与基因3型的欧洲株和中国株同源性为80.8%~82.5%,与基因4型的匈牙利株和中国台湾株同源性为 81.4%~82.6%。ORF2基因进化树分析显示:4株aHEV均属于戊型肝炎B种,形成独立的基因分支,但不属于已报道的4个基因型,而在一个新型独立分支上,表明国内存在一种新基因型的aHEV流行。本研究为我国aHEV的进一步研究提供了基础和依据。


Laboratory Diagnosis of four Cases of Avian Hepatitis E andPhylogenetic Analysis on Its ORF2 Gene

In the study,throughdiagnostic technology of molecular epidemiology,liverand spleen samples were collected from sick chickens with symptoms of hepatitisin four farms in Shandong,Liaoning,Jilin and Shaanxi provinces,then avian hepatitisE virus(aHEV) was detected byRT-PCR and the fragments of aHEV-ORF2 gene were sequenced. At last,the homology was compared by means of MegAlign. The resultsshowed that all samples from the above four farms were positive for aHEV,and the homology between the gene sequences ranged from 84.5% to94.5%. In addition,the homologies of the four isolatedstrains with four already published genotypes were all less than 84.0%,specifically,the homologies with genotypeHEV 1(Australian and Korean strains),HEV 2(American prototype and Americanvirus-free strains),HEV 3(Europeanand Chinese strains),HEV 4(Hungarianand Taiwanese-chinese strains)were 81.6% to 82.8%,80.9% to 82.8%,80.8% to 82.5% and 81.4% to82.6%,respectively. According to the analysis of evolutionarytree of ORF2 gene,the four strains of aHEV allbelonged to B type of hepatitis E,instead of the fourreported genotypes,which formed a new separate genebranch,indicating a new genotype was prevalent inChina. The research would provide some references and foundation for furtherstudy of aHEV in China.

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