Introduction of Digital Polymerase Chain Reaction: Comparison with Real-time Polymerase Chain Reaction

Abstract

Digital polymerase chain reaction (dPCR) is an advanced molecular biology technique that provides a precise and absolute quantitation of target nucleic acids, providing significant improvements over traditional real-time PCR (qPCR) methods. Unlike qPCR, which provides relative quantitation, dPCR offers absolute quantitation by partitioning a sample into tens of thousands of individual reactions, allowing the detection of low-abundance targets amidst a background of abundant wild-type. This technique minimizes the effects of PCR inhibitors and is particularly valuable in applications such as rare sequence detection, rare mutation detection, analysis of copy number variation, liquid biopsy testing, gene expression and RNA analysis, pathogen detection and infectious disease diagnosis, analysis of next-generation sequencing library, and food and environmental monitoring. The ability to detect low-frequency mutations and copy number variations makes dPCR a powerful tool in personalized medicine and cancer research. This review introduces potential markets, history, principles, types, applications, comparison of qPCR and dPCR, and corporations marketing dPCR products. As the technology continues to evolve, dPCR is poised to play a critical role in advancing the understanding of complex biological systems and improving patient outcomes.

Keywords

• absolute quantitation
• digital polymerase chain reaction
• precision
• real-time polymerase chain reaciton
• sensitivity and specificity