PCR检测犬腺病毒方法的建立

Detection of Canine Adenoviruses by Polymerase Chain Reaction

利用人工合成的两条引物,对病犬肝脏和细胞培养物冻融上清中的1、2型犬腺病毒DNA进行多聚酶链式反应（PCR）检测;再将扩增产物进行同位素标记,与纯化病毒DNA进行打点杂交。扩增产物经电泳分析结果表明,其大小与设计的引物间的序列大小基本一致;扩增产物经标记后只与犬腺病毒DNA发生杂交反应,表明其为犬腺病毒特异性核酸片段。

Two primers were synthesized and used to detect Infectious Canine Hepatitis Virus (ICHV) and Canine Laryngotracheitis Virus (CLTV), which were also referred to canine adenovirus type 1 and type 2 respectively, from infected canine liver by Polymerase Chain Reaction(PCR). The amplified sequences were 520 bp and 1 030 bp in size respectively and the former was radiolabelled with isotope and hybridized with ICHV and CLTV DNAs. It was demonstrated that the sequences were specific for canine adenoviruses. It is expected that this technique would be used in the diagnosis of canine adenovirus infection.