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Reduced False Positives and Improved Reporting of Loop-Mediated Isothermal Amplification using Quenched Fluorescent Primers.
Scientific Reports ( IF 3.8 ) Pub Date : 2019-05-14 , DOI: 10.1038/s41598-019-43817-z Patrick Hardinge 1 , James A H Murray 1
Scientific Reports ( IF 3.8 ) Pub Date : 2019-05-14 , DOI: 10.1038/s41598-019-43817-z Patrick Hardinge 1 , James A H Murray 1
Affiliation
Loop-mediated isothermal amplification (LAMP) is increasingly used in molecular diagnostics as an alternative to PCR based methods. There are numerous reported techniques to detect the LAMP amplification including turbidity, bioluminescence and intercalating fluorescent dyes. In this report we show that quenched fluorescent labels on various LAMP primers can be used to quantify and detect target DNA molecules down to single copy numbers. By selecting different fluorophores, this method can be simply multiplexed. Moreover this highly specific LAMP detection technique can reduce the incidence of false positives originating from mispriming events. Attribution of these events to particular primers will help inform and improve LAMP primer design.
中文翻译:
使用淬灭荧光引物减少了假阳性并改进了环介导等温扩增的报告。
环介导的等温扩增 (LAMP) 越来越多地用于分子诊断,作为基于 PCR 的方法的替代方法。有许多报道的技术可以检测 LAMP 扩增,包括浊度、生物发光和嵌入荧光染料。在本报告中,我们展示了各种 LAMP 引物上的淬灭荧光标记可用于量化和检测低至单拷贝数的目标 DNA 分子。通过选择不同的荧光团,这种方法可以简单地进行多路复用。此外,这种高度特异性的 LAMP 检测技术可以减少源自错误启动事件的误报的发生率。将这些事件归因于特定引物将有助于告知和改进 LAMP 引物设计。
更新日期:2019-05-16
中文翻译:
使用淬灭荧光引物减少了假阳性并改进了环介导等温扩增的报告。
环介导的等温扩增 (LAMP) 越来越多地用于分子诊断,作为基于 PCR 的方法的替代方法。有许多报道的技术可以检测 LAMP 扩增,包括浊度、生物发光和嵌入荧光染料。在本报告中,我们展示了各种 LAMP 引物上的淬灭荧光标记可用于量化和检测低至单拷贝数的目标 DNA 分子。通过选择不同的荧光团,这种方法可以简单地进行多路复用。此外,这种高度特异性的 LAMP 检测技术可以减少源自错误启动事件的误报的发生率。将这些事件归因于特定引物将有助于告知和改进 LAMP 引物设计。