Ⅰ群禽腺病毒血清4型广东株的分离鉴定及分子特性  

Isolation, identification and molecular characteristics of group Ⅰ fowl adenovirus serotype 4 from Guangdong, China

作　　者：罗洋洋 招丽婵[2] 李群辉 林瑞庆[1] 翁亚彪[1] LUO Yangyang;ZHAO Lichan;LI Qunhui;LIN Ruiqing;WENG Yabiao(College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China;Guangdong WENS Foodstuff Group Co., Ltd., Yunfu 527400, China)

机构地区：[1]华南农业大学兽医学院,广东广州510642 [2]广东温氏食品集团股份有限公司,广东云浮527400

出　　处：《华南农业大学学报》2017年第6期27-33,共7页Journal of South China Agricultural University

基　　金：国家重点研发计划(2016YFD0501303)

摘　　要：【目的】对广东地区疑似心包积液–肝炎综合征患病鸡进行腺病毒分离鉴定并研究其分子特征。【方法】采集的病料样品,通过鸡胚有限稀释法经SPF鸡胚传代分离纯化病毒,进行致病性试验,并对病毒的hexon、fiber-2、ORF19和ORF29基因进行PCR扩增,分析其分子特征。【结果】hexon基因序列分析显示此次分离的毒株为Ⅰ群禽腺病毒血清4型(命名为WM-XC株),SPF鸡接种分离毒株后,致死率达90%,死亡鸡只出现明显的心包积液、肝炎等变化,肝脏病理切片可观察到明显的嗜酸性包涵体,显示分离毒株具有致病性。fiber-2基因序列同经典毒株ON1和KR5相应序列的相似性达96.1%和97.0%,与2015年以来国内分离的毒株序列相似性则达到100%;ORF29序列与2013年国内分离到的毒株JSJ13和JN13的相应序列相比存在33个碱基的缺失;WM-XC分离株与2015年以来国内分离毒株序列相同,但相比经典株ON1和KR-5,缺失ORF19序列。【结论】此次从广东地区分离到的血清4型Ⅰ群腺病毒,和近年国内其他地区分离的流行毒株一样,其fiber-2、ORF19和ORF29基因序列与经典毒株差异明显。【Objective】To isolate and identify the pathogen causing the inclusion body hepatitis(IBH)and hydropericardium syndrome(HPS)in chicken in Guangdong Province,and study its molecular characteristics.【Method】A virus was isolated from specific pathogen-free(SPF)chicken embryo inoculated with liver suspension from affected chickens.The virus was purified by limiting dilution method and used for pathogenicity test.Hexon,fiber-2,ORF29and ORF19genes were amplified by PCR,and analyzed for molecular characteristics.【Result】Sequence analysis of hexon gene revealed the strain,named as WM-XC,belonged to group I fowl adenovirus serotype4(FAdV-4).This isolated strain was pathogenic based on the evidence that the mortality rate of SPF chicken reached90%after virus inoculation,the hydropericardiumhepatitis syndrome was observed,and acidophilous inclusion body was found in the pathological section of liver.The fiber-2gene sequence of WM-XC had96.1%and97.0%similarities with the corresponding sequences of ON1and KR-5,and had100%similarity with fiber-2genes of fowl adenovirus isolates collected in China since2015.There was a33bp deletion in ORF29gene of WM-XC compared with JSJ13and JN13strains isolated in2013in China.Similar to isolates collected in China since2015,WM-XC lacked ORF19compared with classical FAdV-4strain ON1and KR-5.【Conclusion】The isolated group I FAdV-4strain and other isolates collected in China in recent years were distinct from classical FAdV-4strains in their fiber-2,ORF19and ORF29gene sequences.

关 键 词：禽腺病毒 分离鉴定 分子特性 hexon基因 fiber-2基因 致病性 

分 类 号：S852.659.1[农业科学—基础兽医学]