Accelerated reaction by loop-mediated isothermal amplification using loop primers
Accelerated reaction by loop-mediated isothermal amplification using loop primers
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DOI：
10.1006/mcpr.2002.0415
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发表时间：
2002-06-01
影响因子：
3.3
通讯作者：
Notomi, T
中科院分区：
文献类型：
--
作者：
Nagamine, K;Hase, T;Notomi, T
环介导等温扩增（Loop-mediated isothermal amplification，LAMP）是一种新型的核酸扩增方法，它利用一组4对引物和一种具有链置换活性的DNA聚合酶，在等温条件下高效、特异、快速地扩增DNA。我们已经开发了一种方法，通过使用额外的引物，称为环引物，加速LAMP反应。环引物与茎-环杂交，但与内部引物杂交的环除外，并引发链置换DNA合成。尽管内环引物和环引物都通过环反应，但它们通过不同的机制进行反应。本文介绍的LAMP方法使用环引物，以实现反应时间不到原来的LAMP方法的一半。由于包括检测在内的总分析时间小于1小时，因此这种新方法应有助于遗传分析，包括临床实验室中的遗传诊断。(C)2002爱思唯尔科技有限公司版权所有。
Loop-mediated isothermal amplification (LAMP) is a novel nucleic acid amplification method that amplifies DNA with high specificity, efficiency and rapidity under isothermal conditions using a set of four specially designed primers and a DNA polymerase with strand displacement activity. We have developed a method that accelerates the LAMP reaction by using additional primers, termed loop primers. Loop primers hybridize to the stem-loops, except for the loops that are hybridized by the inner primers, and prime strand displacement DNA synthesis. Although both inner and loop primers react via the loops, they do so by different mechanisms. The LAMP method presented here uses loop primers to achieve reaction times of less than half that of the original LAMP method. Since the total time of analysis including detection is less than 1 h, this new method should facilitate genetic analysis, including genetic diagnosis in the clinical laboratory. (C) 2002 Elsevier Science Ltd. All rights reserved.