Mirabile Alessia, Sangiorgio Giuseppe, Bonacci Paolo Giuseppe, Bivona Dalida, Nicitra Emanuele, Bonomo Carmelo, Bongiorno Dafne, Stefani Stefania, Musso Nicolò
Department of Biomedical and Biotechnological Sciences (BIOMETEC), University of Catania, 95125 Catania, Italy.
U.O.C. Laboratory Analysis Unit, University Hospital Policlinico-San Marco, Via Santa Sofia 78, 95123 Catania, Italy.
Diagnostics (Basel). 2024 Jul 25;14(15):1598. doi: 10.3390/diagnostics14151598.
Digital polymerase chain reaction (dPCR) has emerged as a groundbreaking technology in molecular biology and diagnostics, offering exceptional precision and sensitivity in nucleic acid detection and quantification. This review highlights the core principles and transformative potential of dPCR, particularly in infectious disease diagnostics and environmental surveillance. Emphasizing its evolution from traditional PCR, dPCR provides accurate absolute quantification of target nucleic acids through advanced partitioning techniques. The review addresses the significant impact of dPCR in sepsis diagnosis and management, showcasing its superior sensitivity and specificity in early pathogen detection and identification of drug-resistant genes. Despite its advantages, challenges such as optimization of experimental conditions, standardization of data analysis workflows, and high costs are discussed. Furthermore, we compare various commercially available dPCR platforms, detailing their features and applications in clinical and research settings. Additionally, the review explores dPCR's role in water microbiology, particularly in wastewater surveillance and monitoring of waterborne pathogens, underscoring its importance in public health protection. In conclusion, future prospects of dPCR, including methodological optimization, integration with innovative technologies, and expansion into new sectors like metagenomics, are explored.
数字聚合酶链反应(dPCR)已成为分子生物学和诊断领域的一项开创性技术,在核酸检测和定量方面具有卓越的精度和灵敏度。本综述重点介绍了dPCR的核心原理和变革潜力,特别是在传染病诊断和环境监测方面。强调了它从传统PCR的演变,dPCR通过先进的分区技术对目标核酸进行准确的绝对定量。该综述阐述了dPCR在脓毒症诊断和管理中的重大影响,展示了其在早期病原体检测和耐药基因鉴定方面的卓越灵敏度和特异性。尽管具有优势,但也讨论了诸如实验条件优化、数据分析工作流程标准化以及成本高等挑战。此外,我们比较了各种市售的dPCR平台,详细介绍了它们在临床和研究环境中的特点及应用。此外,该综述探讨了dPCR在水微生物学中的作用,特别是在废水监测和水源性病原体监测方面,强调了其在公共卫生保护中的重要性。总之,探讨了dPCR的未来前景,包括方法优化、与创新技术的整合以及向宏基因组学等新领域的拓展。