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当前位置: 首页 > 文章 > 副猪嗜血杆菌环介导等温扩增快速检测方法的建立 畜牧与兽医 2010,42 (4) 28-32
Position: Home > Articles > Rapid detection for Haemohilus parasuis by loop-mediated isothermal amplification Animal Husbandry & Veterinary Medicine 2010,42 (4) 28-32

副猪嗜血杆菌环介导等温扩增快速检测方法的建立

作  者：
吴诗敏;陈芳艳;陈瑞爱;刘颖;王林川
单  位：
华南农业大学兽医学院;华南农业大学动物科学学院;广东大华农动物保健品股份有限公司
关键词：
副猪嗜血杆菌;LAMP;核酸检测
摘  要：
采用环介导等温扩增技术(loop-mediated isothermal amplification,LAMP)建立了一种快速、灵敏、高度特异的副猪嗜血杆菌(Hps)检测方法。针对Hps16S rRNA基因保守区域设计6条引物,在Bst大片段聚合酶的作用下,对模板DNA进行梯状等温扩增,并且优化LAMP的反应体系,检验其灵敏性与特异性。结果表明:该方法对Hps的最小检测限为0.2pg/μL,灵敏性高于常规PCR方法103倍;完成反应仅需65min。该方法灵敏度高、特异性好,能够作为Hps的快速诊断方法,对于基层和实验室应用均具有良好前景。
译  名：
Rapid detection for Haemohilus parasuis by loop-mediated isothermal amplification
作  者：
WU Shi-min1, CHEN Fang-yan2, CHEN Rui-ai3, LIU Ying3, WANG Lin-chuan1,3(1.College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China;2.College of Animal Science, South China Agricultural University, Guangzhou 510642, China;3.Guangdong Huanong-wens Animal Husbandry Co.Ltd, Xinxing 527400, China)
关键词：
Haemohilus parasuis; LAMP; nucleotide detection
摘  要：
Loop-mediated isothermal amplification (LAMP) assay was developed for the rapid, sensitive and specific detection of Haemohilus parasuis (Hps). A set of six primers were designed based on the 16S rRNA sequence of Hps. LAMP was performed using Bst DNA polymerase (large fragment), and ladder-like DNA fragments were observed on agarose gel electrophoresis. The sensitivity test indicated that the LAMP was 103 times higher than that of PCR. The process takes only 65 minutes. The method described in this study was proved to be a sensitive, specific and rapid way for the detection of Hps. It has a potential application for both laboratorial and pen-side detection of Hps.

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