As a new generation absolute quantitive PCR technique, droplet-based digital PCR (ddPCR) provides the benefits of high sensitivity, precision, and reproducibility. A low-cost, portable, and simple-to-use ddPCR device has emerged as a research hotspot because of the rising demand for point-of-care (POC) detection and clinical diagnosis. Due to its integration and miniaturization properties, microfluidic technology has been used in various ddPCR processes. In this review, we present new theories and methods for the four primary ddPCR processes: droplet production, signal amplification, signal detection, and signal quantification. Integrated microfluidic ddPCR platforms are also introduced. Subsequently, we summarize the applications of ddPCR in liquid biopsy, noninvasive prenatal testing, pathogens detection, and genetically modified organism analysis. At last, the existing challenges and prospects of ddPCR are discussed.

作为新一代绝对定量 PCR 技术，基于液滴的数字 PCR (ddPCR) 具有高灵敏度、精确性和可重复性的优点。由于对床旁 (POC) 检测和临床诊断的需求不断增加，低成本、便携且易于使用的 ddPCR 设备已成为研究热点。由于其集成和小型化特性，微流控技术已被用于各种 ddPCR 过程。在这篇综述中，我们提出了四个主要 ddPCR 过程的新理论和方法：液滴产生、信号放大、信号检测和信号量化。还介绍了集成微流控 ddPCR 平台。随后，我们总结了ddPCR在液体活检、无创产前检测、病原体检测、和转基因生物分析。最后，讨论了ddPCR存在的挑战和前景。