Trouchet Amandine, Gines Guillaume, Benhaim Leonor, Taly Valerie
Centre de Recherche des Cordeliers, UMRS1138, INSERM, CNRS, Sorbonne Université, USPC, Université Paris Cité, Equipe Labellisée Ligue Nationale Contre le Cancer, CNRS SNC 5096, Paris, France.
Laboratoire Gulliver, UMR7083 CNRS/ESPCI Paris-PSL Research University, 10 rue Vauquelin, 75005, Paris, France.
Lab Chip. 2025 Jul 21. doi: 10.1039/d5lc00055f.
Digital PCR (dPCR) is the third generation of PCR technology, after conventional PCR and real-time quantitative PCR. It is based on the partitioning of a PCR mixture supplemented with the sample to analyse into a large number of parallel reactions, so that each partition contains either 0, 1 or a few nucleic acid targets, according to a Poisson distribution. Following PCR amplification, the fraction of positive partitions is extracted from an end-point measurement, allowing the computation of the target concentration. This calibration-free technology presents powerful advantages including high sensitivity, absolute quantification, high accuracy and reproducibility as well as rapid turnaround time and has therefore rapidly spread. Digital PCR offers a wide range of applications in research, clinical diagnostics, and biotechnology. Among the first clinically relevant applications of dPCR was its ability to detect rare genetic mutations within a background of wild-type genes. This breakthrough paved the way to tumour heterogeneity analysis in oncology and enabled liquid biopsy applications, such as the monitoring of treatment response. The scope of dPCR applications has since rapidly extended to include prenatal diagnosis through the detection of aneuploidy or inherited mutations, as well as pathogen identification the detection of virus-specific genes or antibiotic-resistance genes in bacteria. This review focuses on the clinical applications of dPCR, highlighting its advantages over existing technologies and providing an outlook on future developments.
数字PCR（dPCR）是继传统PCR和实时定量PCR之后的第三代PCR技术。它基于将补充了待分析样品的PCR混合物分配到大量平行反应中，从而使每个分区根据泊松分布含有0、1或少数核酸靶标。PCR扩增后，从终点测量中提取阳性分区的比例，从而可以计算靶标浓度。这种无需校准的技术具有强大的优势，包括高灵敏度、绝对定量、高精度和可重复性以及快速周转时间，因此迅速得到推广。数字PCR在研究、临床诊断和生物技术领域有广泛的应用。dPCR最早的临床相关应用之一是能够在野生型基因背景下检测罕见的基因突变。这一突破为肿瘤学中的肿瘤异质性分析铺平了道路，并推动了液体活检应用，如治疗反应监测。此后，dPCR的应用范围迅速扩大，包括通过检测非整倍体或遗传突变进行产前诊断，以及病原体鉴定——检测细菌中的病毒特异性基因或抗生素抗性基因。本综述重点关注dPCR的临床应用，突出其相对于现有技术的优势，并展望未来发展。
