12株肉仔鸡包涵体肝炎病毒的分离和PCR鉴定  

Isolation and PCR identification of 12 strains of inclusion body hepatitis virus from clinical broilers

作　　者：李海英[1] 尹燕博[1] 郭妍妍 胡永钢 许宏伟 王晓红[1] 王守春[1] 

机构地区：[1]青岛农业大学动物科技学院,山东青岛266109 [2]青岛澳兰百特生物工程有限公司,山东青岛266101 [3]青岛市崂山区农林局,山东青岛266061

出　　处：《中国兽医科学》2010年第7期722-727,共6页Chinese Veterinary Science

摘　　要：从临床疑似肉仔鸡包涵体肝炎病例的肝组织中,通过SPF鸡胚连续传代和鸡胚肾细胞培养,分离到12株疑似肉仔鸡包涵体肝炎病毒毒株。通过形态学、致细胞病变和PCR检测等对12株疑似肉仔鸡包涵体肝炎病毒分离物进行了鉴定。结果显示,该病毒为球形、无囊膜、二十面体对称,直径70～80nm;接种鸡胚后鸡胚肝在不同时期表现不同程度的病变;在鸡胚肾细胞(CEK)上可致细胞病变。根据Ⅰ群禽腺病毒CELO株的Hexon保守区基因序列,设计合成了1对引物。用这对引物对12株病毒的核酸模板进行了PCR扩增,结果均特异性地扩增出了与设计片段大小相一致的508bp片段,序列分析表明,分离株确为Ⅰ群禽腺病毒。从病毒形态,鸡胚及细胞病变以及PCR检测结果,证明12株分离物确为肉仔鸡包涵体肝炎病毒。建立的鸡包涵体肝炎快速PCR诊断方法可用于鸡包涵体肝炎的临床快速诊断。The 12 virus strains isolated originally from livers of broiler chickens with inclusion body hepatitis were characterized during chicken embryo kidney（CEK）cell culture.12 isolates of fowl adenovirus groupⅠ were identified based on virus morphology,cytopathogenic effect（CPE）and sequence analysis.The virus particles were non-enveloped,from 70 to 80nm in diameter,and had icosahedrad.Different affections could be observed in embryo eggs during different periods.The virus caused CPE in CEK cells.An avian adenovirus-specific polymerase chain reaction（PCR）was developed.The primers were from the conserved hexon sequence data of the chicken embryo lethal orphan（CELO）avian adenovirus.An avian adenovirus-specific 508bp product was amplified by these primers from hexon.These results confirmed that the 12 isolates were members of chicken inclusion body hepatitis caused by fowl adenovirus groups.The inclusion body hepatitis specific PCR assay was sensitive,specific,rapid and simple,which would be useful for the clinical diagnosis of inclusion body hepatitis virus infection.

关 键 词：鸡 包涵体肝炎病毒 分离 聚合酶链反应 

分 类 号：S855.3[农业科学—临床兽医学] S852.65[农业科学—兽医学]