| 副猪嗜血杆菌环介导等温扩增检测方法的建立 |
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| 引用本文： | 朱杰仪,谭实勇,曾小娜,刘健,刘中勇,朱道中,罗满林.副猪嗜血杆菌环介导等温扩增检测方法的建立[J].中国畜牧兽医,2010,37(2):204-206. |
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| 作者姓名： | 朱杰仪 谭实勇 曾小娜 刘健 刘中勇 朱道中 罗满林 |
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| 作者单位： | （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） |
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| 基金项目： | 广东温氏食品集团有限公司科研基金资助(C080921) |
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| 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。
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| 关 键 词： | 关键词：副猪嗜血杆菌 16S rRNA 环介导等温扩增技术 |
Establishment of Loop-mediated Isothermal Amplification for Detection of Haemophilus parasuis |
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| ZHU Jie-yi,TAN Shi-yong,ZENG Xiao-na,LIU Jian,LIU Zhong-yong,ZHU Dao-zhong,LUO Man-lin.Establishment of Loop-mediated Isothermal Amplification for Detection of Haemophilus parasuis[J].China Animal Husbandry & Veterinary Medicine,2010,37(2):204-206. |
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| Authors: | ZHU Jie-yi TAN Shi-yong ZENG Xiao-na LIU Jian LIU Zhong-yong ZHU Dao-zhong LUO Man-lin |
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| Institution: | （1.Guangdong Key Laboratory of Zoonosis, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China; 2.Guangdong Entry-exit Inspection and Quarantine Bureau, Guangzhou 510623, China） |
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| Abstract: | Abstract：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. |
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| Keywords: | 16S rRNA |
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