蛋鸡J亚群禽白血病的分子生物学诊断

根据J亚群白血病病毒(ALV-J)原型株HPRS-103的序列设计了一对针对外源性ALV-J引物H5和H7,从发生ML病死鸡的肿瘤、骨髓、肝脏、脾脏和输卵管组织中提取DNA作为模板,经PCR扩增得到长度为545bp的片段,对其序列进行测定后,与ALV-J原型株HPRS-103的序列进行了比较,发现其核苷酸同源性为97．4％,所编码氨基酸的同源性为96．1％。该片段含有ALV-J gp85编码基因的部分序列和ALV-J pol基因的部分序列,从分子水平上证实了蛋鸡发生J亚群禽白血病,进一步证明了此前根据病理学观察、免疫组化及免疫荧光诊断的结果。这是首次从分子水平上证明蛋用型鸡发生J亚群禽白血病。

Molecular Diagnosis of Subgroup J Avian Leukosis Virus in Egg-type Chickens

A pair of primers H5/H7 were designed according to the sequence of prototype ALV-J strain HPRS-103. DNAs were extracted from the tumor,bone marrow, oviduct,liver,and spleen from the dead and diseased chickens of various flocks in Northern China.A 545bp fragment was amplified by"touch down"PCR from DNAs of these tissues.The sequence of PCR product was compared with that of ALV-J prototype HPRS-103 strain.The identity of nucleotide was 97.4%,of predicted amino acid was 96.1%.As partial of gp85 encoded gene and 3region of pol gene were included in the amplified fragment,the disease in egg-type chickens could be diagnosed as ALV-J by molecular method exactly.It also demonstrated that the previous diagnostic results of the virus by pathologic method,immunohistochemistry and IFA were accurate.This is the first report at the molecular level to discover ALV-J in egg-type chickens in the world.