| LAMP检测无乳链球菌方法的建立及应用 |
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| 引用本文： | 王永,赵新,景海春,兰青阔,朱珠,程奕.LAMP检测无乳链球菌方法的建立及应用[J].华北农学报,2009,24(5). |
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| 作者姓名： | 王永 赵新 景海春 兰青阔 朱珠 程奕 |
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| 作者单位： | 1. 天津市农业科学院,中心实验室,天津,300381
2. Plant Pathogen Interactions Division,Rothamsted Research,AL5 2JQ UK |
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| 基金项目： | 科技部国际合作项目，天津市农业科学院院偿基金项目 |
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| 摘 要： | 以无乳链球菌纤连蛋白fbs基因为主要研究对象,采取环介导等温扩增技术(Loop-Mediated Isothermal Amplification,LAMP),针对fbs基因的6个区域设计4条特异性引物,利用一种链置换DNA聚合酶(Bst DNA polymerase)在63℃保温1 h,通过荧光显色即可完成对无乳链球菌的检测工作.结果显示,LAMP方法能够特异性检测fbs基因,其检测灵敏度是常规PCR方法的100倍,并与实时荧光定量PCR方法相当.所建立的针对无乳链球菌fbs基因的LAMP检测方法具有高度的特异性及稳定性,结果可靠,非常适合无乳链球菌的快速检测.
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| 关 键 词： | 环介导等温扩增技术 无乳链球菌 检测 |
Development and Application of Loop-Mediated Isothermal Amplification for Detection of Streptococcus agalactiae |
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| WANG Yong,ZHAO Xin,JING Hai-chun,LAN Qing-kuo,ZHU Zhu,CHENG Yi.Development and Application of Loop-Mediated Isothermal Amplification for Detection of Streptococcus agalactiae[J].Acta Agriculturae Boreali-Sinica,2009,24(5). |
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| Authors: | WANG Yong ZHAO Xin JING Hai-chun LAN Qing-kuo ZHU Zhu CHENG Yi |
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| Abstract: | To develop a technique for detecting the fbsB gene of Streptococcus agalactiae using a novel DNA amplification method designated Loop-Mediated Isothermal Amplification (LAMP).The detection assay is based on the loop-mediated isothermal amplification (LAMP) reaction.The fbsB gene was amplified by a set of four specially primers that recognize six distinct sequences of the target.The amplification can be obtained in 1 h by incubating all of the reagents in a single tube with Bst DNA polymerase at 63℃.Results showed that the developed LAMP assay demonstrated an exceptionally higher than the conventional PCR.This assay results was found to be 100 times more sensitive than the PCR assay,and consistent with the results of real-time PCR.Our results clearly demonstrate that the LAMP-based assay is a sensitive and reliable means for the detection of Streptococcus agalactiae. |
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| Keywords: | fbsB Loop-mediated isothermal amplification(LAMP) Streptococcus agalactiae fbsB Detection |
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