Real-time turbidimetry of LAMP reaction for quantifying template DNA
Real-time turbidimetry of LAMP reaction for quantifying template DNA
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DOI：
10.1016/j.jbbm.2003.12.005
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发表时间：
2004-05-31
影响因子：
--
通讯作者：
Notomi, T
中科院分区：
文献类型：
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作者：
Mori, Y;Kitao, M;Notomi, T
环介导的等温扩增(LAMP)是一种核酸扩增方法，它允许在短时间内合成大量的DNA，并且具有很高的特异性。随着LAMP反应的进行，反应副产物焦磷酸盐离子与镁离子结合，形成白色焦磷酸镁沉淀物。我们设计了一种能够在保持恒温的情况下同时测量多个样品的浊度的装置，以实时测量LAMP反应的浊度变化。LAMP反应溶液的浊度超过给定值所需的时间(T_1)取决于初始模板DNA的量。也就是说，T-t与初始模板DNA量对数图在模板DNA的2×10(3)拷贝(0.01pg/管)到2×10(9)拷贝(100 ng/管)之间呈线性关系。这些结果表明，LAMP反应的实时浊度测量允许使用本研究报告的一种简单设备对样品中存在的微量核酸进行定量分析，在很大范围内具有高精度。(C)2003爱思唯尔B.V.保留所有权利。
Loop-mediated isothermal amplification (LAMP) is a nucleic acid amplification method that allows the synthesis of large amounts of DNA in a short period of time with high specificity. As the LAMP reaction progresses, the reaction by-product pyrophosphate ions bind to magnesium ions and form a white precipitate of magnesium pyrophosphate. We designed an apparatus capable of measuring the turbidity of multiple samples simultaneously while maintaining constant temperature to conduct real-time measurements of the changes in the turbidity of LAMP reactions. The time (T,) required for the turbidity of the LAMP reaction solution to exceed a given value was dependent on the quantity of the initial template DNA. That is, a graph with the plot of T-t versus the log of the amount of initial template DNA was linear from 2 x 10(3) copies (0.01 pg/tube) to 2 x 10(9) copies (100 ng/tube) of template DNA. These results indicate that real-time turbidity measurements of the LAMP reaction permit the quantitative analysis of minute amounts of nucleic acids present in a sample, with a high precision over a wide range, using a simple apparatus reported in this study. (C) 2003 Elsevier B.V. All rights reserved.