摘要
为了解山西地区4型禽腺病毒(FAdV-4)的遗传进化情况及其致病性,为山西省FAdV-4的疫苗研发和防控策略制定提供理论依据,收集了山西中部地区太谷、祁县、介休、临汾、长治、交城、柳林等养鸡场11份疑似心包积水-肝炎综合征病鸡的肝脏组织样本,采用PCR方法进行FAdV-4检测,对阳性样品进行FAdV-4的Hexon和Fiber-2蛋白的全基因序列扩增,将处理后的阳性病料接种于鸡肝癌细胞(LMH)进行病毒分离和PCR鉴定,测定分离毒株的TCID50,研究病毒对SPF鸡胚和SPF鸡的致病性。结果表明,11份样品的FAdV-4检测均为阳性,经比对,11份样品中FAdV-4的Hexon和Fiber-2基因序列相同,与GenBank中不同FAdV分离株的同源性分别为67.7%~100%和43.2%~100%,其中,与GenBank中的基因C型FAdV-4毒株处于同一个分支,同源性分别为98.4%~100%和95.9%~100%,说明11份疑似样品均存在FAdV-4感染,且为同一毒株。分离获得的1株FAdV-4(命名为SX2020)可在LMH细胞中良好复制并产生明显的细胞病变,对SPF鸡胚及SPF鸡均有致病性,感染后出现FAdV-4典型病变,并且病毒可以在感染鸡体内持续向外排毒达21~28 d。
In order to understand genetic evolution and pathogenicity of avian adenovirus type 4(FAdV-4)in Shanxi province,provide theoretical foundation for research and development of vaccines and prevention-control strategy of FAdV-4 in Shanxi province,in this study,11 liver samples of chicken suspected of hydropericardium hepatitis syndrome were collected from Taigu,Qixian,Jiexiu,Linfen,Changzhi,Jiaocheng and Liulin in central Shanxi province,and FAdV-4 were detected by PCR method.The Hexon of FAdV-4 and gene sequence amplification of Fiber-2 protein of the positive samples were conducted.The positive samples treated were inoculated into chicken hepatoma cells(LMH)for virus isolation and PCR identification.The TCID50 of the isolated strain was determined,and pathogenicity of the isolated virus to SPF chicken embryos and SPF chicken was studied.The results showed that all 11 samples were positive for FAdV-4,and the sequences of Hexon and Fiber-2 genes of FAdV-4 in 11 samples were the same.The homology of Hexon and Fiber-2 gene with different FAdV isolates in GenBank was67.7%-100%and 43.2%-100%.They were in the same branch with FAdV-4 C strain in GenBank,and the homology with FAdV-4 was 98.4%-100%and 95.9%-100%,indicating that all the 11 suspected samples were infected by FAdV-4,and they were the same strain.One isolated FAdV-4 virus(named SX2020)could replicate well in LMH cells and produce obvious cytopathic changes(CPE).The virus was pathogenic to SPF chicken embryos and SPF chicken,and the typical lesions of FAdV-4 appeared after infection.The virus could continuously excrete outward in infected chicken for up to 21-28 d.
作者
刘璐
王晨
张丹
高宇
程宇
谭世明
梁立滨
马海利
LIU Lu;WANG Chen;ZHANG Dan;GAO Yu;CHENG Yu;TAN Shiming;LIANG Libin;MA Haili(College of Veterinary Medicine,Shanxi Agricultural University,Taigu 030801,China)
出处
《山西农业科学》
2022年第4期591-597,共7页
Journal of Shanxi Agricultural Sciences
基金
山西省重点研发计划重点项目(201703D211001)。
关键词
4型禽腺病毒
分离与鉴定
遗传进化
致病性
Avian adenovirus type 4(FAdv4)
isolation and identification
genetic evolution
pathogenicity
