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PCR

PCR

PCR - Polymerase Chain Reaction

PCR - Polymerase Chain Reaction

Real Time RT-PCR

Real Time RT-PCR

Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...

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Application of Digital Polymerase Chain Reaction (dPCR) in Non-Invasive Prenatal Testing (NIPT).

Ying Guo^1,2, Pimlak Charoenkwan^3,4, Kuntharee Traisrisilp¹

¹Department of Obstetrics and Gynaecology, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand.

|March 28, 2025

Summary

Digital PCR (dPCR) offers a precise and cost-effective alternative for non-invasive prenatal testing (NIPT), potentially matching Next-Generation Sequencing (NGS) accuracy. This method shows promise for wider accessibility in prenatal genetic screening.

Keywords:

chromosomal aneuploidy chromosomal microdeletions and microduplications digital PCR (dPCR)monogenic disease non-invasive prenatal testing (NIPT)

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Area of Science:

Biotechnology
Genetics
Maternal-Fetal Medicine

Background:

Rising incidence of genetic disorders necessitates advanced prenatal testing.
Non-invasive prenatal testing (NIPT) aims to reduce invasive procedures.
Cell-free fetal DNA (cffDNA) analysis is central to NIPT.

Purpose of the Study:

To review digital PCR (dPCR) applications in NIPT.
To compare dPCR with Next-Generation Sequencing (NGS) for prenatal testing.
To assess dPCR's potential to overcome NIPT challenges.

Main Methods:

Review of current dPCR applications in NIPT.
Comparative analysis of dPCR and NGS performance metrics.
Evaluation of dPCR's robustness against interfering factors in cffDNA analysis.

Main Results:

dPCR demonstrates precision in detecting low-abundance cffDNA targets.
dPCR is robust against maternal DNA contamination and DNA degradation.
dPCR offers comparable accuracy to NGS at lower cost and complexity.

Conclusions:

dPCR presents a viable and potentially superior alternative to NGS for NIPT.
The precision and cost-effectiveness of dPCR enhance accessibility for prenatal genetic screening.
dPCR is a promising technology for improving maternal-fetal medicine outcomes.