副猪嗜血杆菌环介导等温扩增检测方法的建立  

Establishment of Loop-mediated Isothermal Amplification for Detection of Haemophilus parasuis

作　　者：朱杰仪[1] 谭实勇[1] 曾小娜[1] 刘健[1] 刘中勇[2] 朱道中[2] 罗满林[1] 

机构地区：[1]华南农业大学兽医学院广东省人畜共患病重点实验室,广州510642 [2]广东出入境检验检疫局,广州510623

出　　处：《中国畜牧兽医》2010年第2期204-207,共4页China Animal Husbandry & Veterinary Medicine

基　　金：广东温氏食品集团有限公司科研基金资助(C080921)

摘　　要：本研究旨在建立检测副猪嗜血杆菌(Hps)的新方法。根据GenBank上发表的不同血清型Hps的16S rRNA序列,找出高度保守区域,再针对该区域设计了4条特异性引物,建立了Hps环介导等温扩增法(LAMP)。试验结果表明,LAMP方法在恒温(63℃)条件下特异性强,比PCR方法敏感100倍,鉴于目前副猪嗜血杆菌分离培养和鉴定的复杂性,LAMP操作简单、成本低,整个扩增反应在1 h内即可完成,为检测Hps提供了一个快速简便的分子生物学方法。Based on a series of 16S rRNA sequences of Haemophilus parasuis（Hps） published in GenBank,a conservative region was found.Four specific primers targeting the 6 distinct sequences of the conservative region were designed.A loop-mediated isothermal amplification（LAMP） for the rapid detection of Hps was established by using those specific primers.The results show that LAMP has a high specificity,and is more sensitive than PCR method under 63 ℃.Compared with the complex isolation and identification methods of Hps,LAMP is simple,low cost,and could be completed in 1 h.

关 键 词：副猪嗜血杆菌 16S RRNA 环介导等温扩增技术 

分 类 号：S852.65[农业科学—基础兽医学]