数字聚合酶链式反应在检验诊断中的应用进展

Advances in the application of digital polymerase chain reaction in laboratory diagnosis

数字聚合酶链式反应(dPCR)是继实时定量聚合酶链反应之后发展起来的不依赖于标准曲线的高灵敏度和高特异性核酸绝对定量分析技术。dPCR的PCR反应体系被分成大量独立的反应分子池,在其进行PCR扩增时,核酸拷贝数根据泊松分布校正,最终,实现低浓度样品的绝对定量分析。近年来,基于微流控技术的发展数字聚合酶链式反应检测技术准确性和特异性明显提高。本文详细介绍了数字聚合酶链反应技术的原理,阐述了其在临床疾病诊断方面的应用概况和技术发展。

Digital Polymerase chain reaction(dPCR)is an absolute quantitative nucleic acid analysis technique with a high sensitivity and specificity,and independent of standard curve developed after realtime quantitative polymerase chain reaction.The PCR reaction system of dPCR is divided into many independent reaction molecular pools,where PCR amplification is performed,and the nucleic acid copy number is corrected according to Poisson distribution.Finally,the absolute quantitative analysis of low concentration samples is achieved.In recent years,based on the development of microfluidic technology,the accuracy and specificity of dPCR detection technology have been significantly improved.This paper introduces the principle of dPCR technology in detail,and expounds its application and technology development in clinical disease diagnosis.