Current, Emerging, and Future Applications of Digital PCR in Non-Invasive Prenatal Diagnosis

Abstract

Digital PCR (dPCR) approaches have been developed for the detection of nucleic acids of low abundance, such as cell-free DNA, and represent an attractive and sensitive alternative to conventional methods, particularly in the field of non-invasive prenatal diagnosis (NIPD). In this review, we present the principle of dPCR and its applications in the field of prenatal diagnosis from current and emerging uses, such as fetal gender determination, rhesus blood group D antigen genotyping, or monogenic disorders prenatal testing, to future applications, such as the diagnosis and monitoring of pregnancy-related disorders. We also address considerations for implementation of the method in a clinical laboratory and discuss the competiveness of dPCR over other technologies such as quantitative PCR or massively parallel sequencing.

Evaluation of droplet digital PCR for quantification of SARS-CoV-2 Virus in discharged COVID-19 patients

“…Although quantitative real-time PCR (qPCR) detection of nucleic acids from the virus has become a standard method for diagnosis of SARS-CoV-2 infection, this approach is limited by an unsatisfactory amplification curve, which can lead to the occurrence of false negatives [3][4][5]. The high false negative rate and the lack of availability of qPCR assays for discharged patients during the outbreak restricted prompt diagnosis of infected patients [6,7]. In addition, the qPCR method does not provide information about viral load, which would enable evaluation of disease progression and prognosis.…”

An image-to-answer algorithm for fully automated digital PCR image processing

“…Unfortunately, there is a chance that multiple targets can land in the same partitions. 45 In this case, the data extraction described above using two Gauss distribution functions would not yield correct results. The probability P ( k ) of having k targets in a partition is given by Poisson distribution: 23 where λ is the average cn in each partition.…”

Noninvasive diagnostics of fetal KEL*01.01 allele from maternal plasma of immunized women using digital PCR protocols

“…[12][13][14] Moreover, the technique is a robust and inexpensive alternative to other solutions such as NGS, also known to detect fetal fraction. 15 The aim of our work was to evaluate different digital PCR protocols for NIPT of the fetal KEL*01.01 genotype offered to immunized pregnant women with anti-K antibodies. It is the first work comparing the usefulness of digital PCR protocols used for NIPT of fetal SNV-based alleles.…”

“…The digital PCR technique is known to specifically detect minor fractions with single nucleotide mutations hidden in the highly concentrated material of the opposite allele such as the plasma DNA of pregnant women due to the dividing of the reaction into thousands of microcells 12–14 . Moreover, the technique is a robust and inexpensive alternative to other solutions such as NGS, also known to detect fetal fraction 15 . The aim of our work was to evaluate different digital PCR protocols for NIPT of the fetal KEL*01.01 genotype offered to immunized pregnant women with anti‐K antibodies.…”