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

作　　者：刘涛[1] LIU Tao(National Health Commission’s Key Laboratory of Critical Care Medicine,Tianjin First Central Hospital Affiliated to Nankai University,Tianjin 300192,China)

机构地区：[1]南开大学附属第一中心医院国家卫生健康委员会危重病急救医学重点实验室,天津300192

出　　处：《生命的化学》2022年第4期781-787,共7页Chemistry of Life

基　　金：国家自然科学基金项目(81402322);南开大学附属第一中心医院科技基金春苗项目(2020CM01)。

摘　　要：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.

关 键 词：微RNA 逆转录-定量PCR 实时定量PCR 

分 类 号：Q503[生物学—生物化学]