Quantitative and Digital PCR in Biotechnology: Methodological Considerations and Applications
Kitapta bölümler > Diğer · 2026
Yazarlar: Gulnar Ahmadova, Memiş Bilgici
Principles and Applications of Polymerase Chain Reaction [Working Title], Diğer, IntechOpen, B, Uluslararası, 2026, ,
DOI: 10.5772/intechopen.1016323
Özet
Quantitative polymerase chain reaction (qPCR) and digital polymerase chain reaction (dPCR) represent the two primary pillars of nucleic acid quantification in modern biotechnology and clinical molecular diagnostics. While both technologies utilize the fundamental principles of thermal cycling and fluorescence detection, they differ significantly in their methodological approach, quantification kinetics, and data interpretation. This chapter provides a comprehensive review of their core mechanisms, comparative analytical performance, and critical considerations essential for robust assay design, validation, and laboratory implementation. qPCR remains widely implemented due to its broad dynamic range, high throughput, and cost-effectiveness; however, its reliance on relative quantification demands rigorous calibration strategies and remains highly sensitive to variations in reaction efficiency. Conversely, dPCR achieves absolute target quantification without external standard curves by partitioning the sample into thousands of discrete microreactors. By applying Poisson statistical analysis to the ratio of positive and negative partitions, dPCR offers distinct advantages for rare-target detection, copy number variation (CNV) analysis, and measurements in inhibitor-rich samples. Beyond classical workflows, this chapter explores next-generation modalities, including novel DNA-aptamer-based “light-up” dyes, absolute N calculations and the integration of artificial intelligence (AI) using architectures like XGBoost for autonomous data processing. Looking toward the future, the convergence of these platforms with AI-driven diagnostic pipelines is set to revolutionize personalized medicine by enabling high-precision clinical decision-making. Ultimately, qPCR and dPCR should be viewed as complementary rather than competing technologies. The selection of the most appropriate platform must be strategically determined by the specific analytical objective, required sensitivity, tissue-specific matrix constraints, workflow velocity, and budget discipline.