数字聚合酶链反应及其在感染性疾病中的应用

数字聚合酶链反应(polymerase chain reaction,PCR)采用与定量PCR相同的荧光化学原理和不同的数学原理来实现对靶标核酸序列的绝对定量,其摒弃了对外部参照的依赖,同时具有更高的数据精密度,提高了重复性和再现性。数字PCR的应用涵盖生命科学众多领域,特别是在医学检验领域,其对疾病相关核酸分子标记的准确分析,为疾病的早期诊断、进展监测、疗效评估提供了动态量化指标。数字PCR的出现将推动基于核酸扩增技术的分子生物学检测迈入精准定量阶段。本文就数字PCR尤其是微滴式数字PCR在感染性疾病中的应用进展及前沿进行综述。

Principle of digital polymerase chain reaction and its application in infectious diseases

Digital polymerase chain reaction (PCR) is an emerging technology for quantitative analysis of nucleic acids, employing the same fluorescence chemistry as quantitative PCR (qPCR) but different mathematical principles to achieve absolute quantification of target nucleic acid sequences, without the reliance on external references such as standard curve. This technology is both conceptually simple and extremely robust in terms of assay performance, including increased sensitivity, precision, reproducibility and repeatability. Digital PCR technology has been widely used in life science research and applications. In the field of medical testing, digital PCR provides accurate quantification of disease-related nucleic acid biomarkers, enabling early diagnosis of diseases, monitoring of disease progression and evaluation of therapeutic efficacy as well. The emergence of digital PCR will push the medical testing into the stage of precise quantification. Here we review the progress and cutting edge of digital PCR technology, especially droplet digital PCR technology in the field of infectious diseases.