本研究通過分子流行病學(xué)診斷技術(shù)，分別從山東、遼寧、吉林、陜西4個地區(qū)表現(xiàn)肝炎癥狀的病雞中，采集肝臟和脾臟樣品，利用RT-PCR方法，檢測禽戊型肝炎病毒（aHEV），并對aHEV-ORF2 基因片段進行測序，利用MegAlign進行同源性比較。結(jié)果顯示：4個雞場的病雞樣品均為aHEV 陽性，aHEV-ORF2基因序列間同源性為84.5%~94.5%，與已報道的4種基因型同源性均小于84.0%，其中與基因1型的澳大利亞株和韓國株同源性為81.6%~82.8%，與基因2型的美國原型株和美國無毒株同源性為80.9%~82.8%，與基因3型的歐洲株和中國株同源性為80.8%~82.5%，與基因4型的匈牙利株和中國臺灣株同源性為 81.4%~82.6%。ORF2基因進化樹分析顯示：4株aHEV均屬于戊型肝炎B種，形成獨立的基因分支，但不屬于已報道的4個基因型，而在一個新型獨立分支上，表明國內(nèi)存在一種新基因型的aHEV流行。本研究為我國aHEV的進一步研究提供了基礎(chǔ)和依據(jù)。
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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