Gleerup David, Trypsteen Wim, Fraley Stephanie I, De Spiegelaere Ward
Laboratory of Veterinary Morphology, Faculty of Veterinary Medicine, Ghent University, Campus Merelbeke, Salisburylaan 133, 9820, Merelbeke, Belgium.
Ghent University Digital PCR Consortium, Ghent University, Ghent, Belgium.
Mol Diagn Ther. 2025 Jan;29(1):43-54. doi: 10.1007/s40291-024-00751-9. Epub 2024 Nov 2.
Digital PCR (dPCR) has been used in the field of virology since its inception. Technological innovations in microfluidics more than a decade ago caused a sharp increase in its use. There is an emerging consensus that dPCR now outperforms quantitative PCR (qPCR) in the basic parameters such as precision, sensitivity, accuracy, repeatability and resistance to inhibitors. These strengths have led to several current applications in quantification, mutation detection and environmental DNA and RNA samples. In high throughput scenarios, such as the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic, the cost and throughput still significantly hampered the adaption of dPCR. There is much unexplored potential within the multiplexing capabilities of dPCR. This will allow simultaneous multi-target quantification and can also partially alleviate the throughput and cost drawback. In this review, we discuss the strengths and weaknesses of dPCR with a focus on virology applications and we discuss future applications. Finally, we discuss recent evolutions of the technology in the form of real-time dPCR and digital high-resolution melting.
数字PCR（dPCR）自问世以来就一直应用于病毒学领域。十多年前微流控技术的创新使其应用急剧增加。目前已逐渐形成一种共识，即dPCR在诸如精密度、灵敏度、准确性、重复性和抗抑制剂能力等基本参数方面优于定量PCR（qPCR）。这些优势促成了dPCR目前在定量分析、突变检测以及环境DNA和RNA样本检测等方面的多种应用。在高通量场景下，例如严重急性呼吸综合征冠状病毒2（SARS-CoV-2）大流行期间，成本和通量仍然严重阻碍了dPCR的应用。dPCR的多重检测能力还有很大未被开发的潜力。这将实现同时多靶点定量分析，也能部分缓解通量和成本方面的劣势。在本综述中，我们将讨论dPCR的优缺点，重点关注病毒学应用，并探讨其未来应用。最后，我们将讨论该技术以实时dPCR和数字高分辨率熔解形式呈现的最新进展。
