禽白血病病毒斑点杂交检测方法的建立  

Establishment of dot-blot hybridization assay for detection of avian leukosis virus

作　　者：周刚[1] 牛成明 司昌德[1] 韩凌霞[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2010年第1期53-55,70,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：中央级公益性科研院所基本科研业务费项目(2009-08)

摘　　要：为建立禽白血病病毒(ALV)斑点杂交检测方法,本研究采用RT-PCR技术扩增ALV群特异性p27抗原基因的部分片段,并以纯化的p27PCR产物为模板,合成地高辛标记探针,以此建立了ALV的斑点杂交检测方法。用该方法对4份疑似感染ALV的现地病鸡组织样品和18枚鸡胚进行检测,结果表明所有样品均为阳性,而且与PCR检测结果的符合率达到100%。该方法具有良好的特异性和敏感性,适应于ALV临床大规模检测。The gene of avian leukosis virus p27 group specific antigen was amplified by RT-PCR and labeled with Digoxigenin-labelled as probe. Dot-blot hybridization was developed and used to detect 4 clinical samples and 18 chicken embryos of suspected ALV infection. The result showed that all samples were detected positive by dot-blot, which was consistent with PCR re- suits.

关 键 词：禽白血病病毒 杂交检测 斑点 RT-PCR技术 地高辛标记探针 PCR产物 ALV 抗原基因 

分 类 号：S852.65[农业科学—基础兽医学] Q78[农业科学—兽医学]