Abdullah Al-Maskri Abdu Ahmed, Ye Jiawei, Talap Jadera, Hu Haihong, Sun Lianli, Yu Lushan, Cai Sheng, Zeng Su
Institute of Drug Metabolism and Pharmaceutical Analysis, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research, College of Pharmaceutical Science, Zhejiang University, Hangzhou, Zhejiang, 310058, China.
Institute of Drug Metabolism and Pharmaceutical Analysis, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research, College of Pharmaceutical Science, Zhejiang University, Hangzhou, Zhejiang, 310058, China.
Anal Chim Acta. 2020 Aug 22;1126:1-6. doi: 10.1016/j.aca.2020.06.007. Epub 2020 Jun 17.
A novel reverse transcription-based loop-mediated isothermal amplification (LAMP) strategy for miRNA detection has been developed. This method consists of two stem-loop probes inspired by the dumbbell-shaped amplicons and inner primers used in conventional LAMP reactions. Termed "terminal hairpin formation and self-priming" (THSP), this reaction incorporates phosphorothioated (PS) modifications to achieve DNA folding and extension without primers. The final signal is monitored by a sequence-specific detection probe, which minimizes the background noise. We suggest that our rapid, facile, and reliable LAMP method will be a promising candidate for detecting miRNA in biomedical applications.
一种用于检测miRNA的基于逆转录的新型环介导等温扩增（LAMP）策略已被开发出来。该方法由两个受传统LAMP反应中哑铃形扩增子和内部引物启发的茎环探针组成。这种反应被称为“末端发夹形成和自引发”（THSP），它采用硫代磷酸化（PS）修饰来实现无需引物的DNA折叠和延伸。最终信号由序列特异性检测探针监测，从而将背景噪声降至最低。我们认为，我们快速、简便且可靠的LAMP方法将成为生物医学应用中检测miRNA的一个有前途的候选方法。
