Laboratory of Pharmaceutical Analysis and Drug Metabolism, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research , Zhejiang University , Hangzhou , Zhejiang 310058 , China.
Division of Biotechnology , College of Life Sciences and Biotechnology, Korea University , Seoul 02841 , Republic of Korea.
Anal Chem. 2018 Jul 17;90(14):8290-8294. doi: 10.1021/acs.analchem.8b02062. Epub 2018 Jul 5.
Loop-mediated isothermal amplification (LAMP) is an extremely powerful tool for the detection of nucleic acids with high sensitivity and specificity. However, LAMP shows optimal performance at around 65 °C, which limits applications in point-of-care-testing (POCT). Here, we have developed a version of LAMP that uses phosphorothioated primers (PS-LAMP) to enable more efficient hairpin formation and extension at the termini of growing concatamers, and that therefore works at much lower temperatures. By including additional factors such as chaotropes (urea) and single-stranded DNA binding protein (SSB), the sensitivities and selectivities for amplicon detection with PS-LAMP at 40 °C were comparable with a regular LAMP reaction at 65 °C.
环介导等温扩增 (LAMP) 是一种非常强大的工具，可用于高灵敏度和特异性的核酸检测。然而，LAMP 在 65°C 左右表现出最佳性能，这限制了其在即时检测 (POCT) 中的应用。在这里，我们开发了一种使用硫代磷酸化引物 (PS-LAMP) 的 LAMP 版本，该版本能够在生长的连接物末端更有效地形成发夹并进行延伸，因此可以在更低的温度下工作。通过包括额外的因素，如变构剂 (尿素) 和单链 DNA 结合蛋白 (SSB)，在 40°C 下使用 PS-LAMP 进行扩增子检测的灵敏度和选择性与在 65°C 下进行常规 LAMP 反应相当。
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