Digital PCR in noninvasive prenatal testing: Analytical principles, clinical utilities, and future integration

Digital PCR in noninvasive prenatal testing: Analytical principles, clinical utilities, and future integration

Článek WoS

Digital PCR (dPCR) enables absolute nucleic acid quantification through partition-based amplification and has emerged as a robust analytical platform for non-invasive prenatal testing (NIPT). Direct molecular counting, independent of calibration curves and amplification efficiency, enables precise detection of chromosomal aneuploidies, rare paternal or de novo variants, and microdeletions at very low fetal DNA fractions where sequencing-based screening loses statistical power. High partition numbers, strong reproducibility, and minimal computational requirements distinguish dPCR from qPCR and next-generation sequencing (NGS), positioning it as a reliable confirmatory method for sequencing-derived findings. Analytical advances, including improved extraction efficiency, enhanced partition stability, expanded multiplexing strategies, and duplex ratio assays, further improve performance in complex plasma samples. Hybrid workflows integrating NGS-based genomewide discovery with dPCR-based quantitative validation reduce false positives and enhance diagnostic confidence. Digital PCR thus complements sequencing and supports a precise and scalable framework for nextgeneration prenatal diagnostics.

Digital PCR (dPCR) enables absolute nucleic acid quantification through partition-based amplification and has emerged as a robust analytical platform for non-invasive prenatal testing (NIPT). Direct molecular counting, independent of calibration curves and amplification efficiency, enables precise detection of chromosomal aneuploidies, rare paternal or de novo variants, and microdeletions at very low fetal DNA fractions where sequencing-based screening loses statistical power. High partition numbers, strong reproducibility, and minimal computational requirements distinguish dPCR from qPCR and next-generation sequencing (NGS), positioning it as a reliable confirmatory method for sequencing-derived findings. Analytical advances, including improved extraction efficiency, enhanced partition stability, expanded multiplexing strategies, and duplex ratio assays, further improve performance in complex plasma samples. Hybrid workflows integrating NGS-based genomewide discovery with dPCR-based quantitative validation reduce false positives and enhance diagnostic confidence. Digital PCR thus complements sequencing and supports a precise and scalable framework for nextgeneration prenatal diagnostics.

Droplet digital polymerase chain reaction, Noninvasive prenatal testing, Cell-free fetal DNA (cffDNA), Next-generation sequencing

Droplet digital polymerase chain reaction, Noninvasive prenatal testing, Cell-free fetal DNA (cffDNA), Next-generation sequencing

ZENG, W.; LUO, W.; ZHOU, J.; WANG, B.; QIN, H.; HE, Y.; GABLECH, I.; KORABECNA, M.; NEUŽIL, P.

2026

01.04.2026

Elsevier

TRAC-TRENDS IN ANALYTICAL CHEMISTRY

197

4

Spojené království Velké Británie a Severního Irska

14

https://www.sciencedirect.com/science/article/pii/S0165993626000506