基于RT-qPCR技术的microRNA表达水平检测方法

Research progress of microRNA expression level detection methods based on RT-qPCR

微RNA(microRNA,miRNA)是一类细胞内源性的小分子非编码RNA,在多种生理和病理过程中发挥基因调控作用。MiRNA的表达水平定量分析是其功能研究中的重要过程,但miRNA的成熟体序列较短,因而其逆转录-定量聚合酶链式反应(reverse transcription quantitative polymerase chain reaction,RT-qPCR)需采取与长链基因不同的特殊策略。现有miRNA的RT-qPCR方法可分为4种类型,包括茎环引物法、加尾法、直接逆转录法和其他方法。在PCR过程中对荧光信号的监测主要有探针法和染料法两种方式。本文对miRNA的RT-qPCR各种方法及其主要原理进行综述。

MicroRNAs(miRNAs) are a class of endogenous small non-coding RNAs that mediate regulation of gene expression in several physiological and pathological processes.Quantitative analysis of miRNA expression level is a key procedure in miRNA functional study.Due to the short length of miRNAs,special strategies as distinguished from those used for long-chain genes should be introduced in miRNA reverse transcription quantitative polymerase chain reaction(RT-qPCR) analysis.Four strategies are currently used in miRNA RT-qPCR including stem-loop primer based method,end-tailing based method,direct reverse transcription method,and other methods.Monitoring of fluorescent signal during PCR includes probe based and nucleic acid dye based methods.In this review,we summarize the strategies of miRNA RT-qPCR and the principle.