化学
环介导等温扩增
逆转录环介导等温扩增
放大器
小RNA
逆转录酶
哑铃
启动(农业)
寡核苷酸
多路复用
逆转录聚合酶链式反应
核糖核酸
分子生物学
DNA
计算生物学
生物物理学
癌症研究
聚合酶链反应
信使核糖核酸
基因
生物化学
生物信息学
生物
生理学
发芽
植物
作者
Abdu Ahmed Abdullah AL-maskri,Jiawei Ye,Jadera Talap,Haihong Hu,Lianli Sun,Lushan Yu,Sheng Cai,Su Zeng
标识
DOI:10.1016/j.aca.2020.06.007
摘要
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. 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. The final signal is monitored by a toehold-mediated strand exchange reaction (one-step strand displacement probe), which is able to minimize background noise. We hope this strategy can provide a highly selective and sensitive platform for miRNA detection. • A novel method of reverse transcription-based miRNA detection has been introduced. • It is based on the dumbbell-shaped amplicons and inner primers used in LAMP. • It has been termed as “terminal hairpin formation and self-priming” (THSP) reaction. • Self-folding efficiency was improved by phosphorothioated modification.
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