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dPCR: A Technology Review

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Quan, Phenix-Lan, Sauzade, Martin, Brouzes, Eric·2018·Sensors
DOI 10.3390/s18041271PMC PMC5948698
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Abstract

Digital Polymerase Chain Reaction (dPCR) is a novel method for the absolute quantification of target nucleic acids. Quantification by dPCR hinges on the fact that the random distribution of molecules in many partitions follows a Poisson distribution. Each partition acts as an individual PCR microreactor and partitions containing amplified target sequences are detected by fluorescence. The proportion of PCR-positive partitions suffices to determine the concentration of the target sequence without a need for calibration. Advances in microfluidics enabled the current revolution of digital quantification by providing efficient partitioning methods. In this review, we compare the fundamental concepts behind the quantification of nucleic acids by dPCR and quantitative real-time PCR (qPCR). We detail the underlying statistics of dPCR and explain how it defines its precision and performance metrics. We review the different microfluidic digital PCR formats, present their underlying physical principles, and analyze the technological evolution of dPCR platforms. We present the novel multiplexing strategies enabled by dPCR and examine how isothermal amplification could be an alternative to PCR in digital assays. Finally, we determine whether the theoretical advantages of dPCR over qPCR hold true by perusing studies that directly compare assays implemented with both methods.

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  • dPCR requires low DNA quantities.

    “It requires low DNA quantities and provides exquisite sensitivity for absolute quantification of target nucleic acids [54,55,56].”

    From Genetic Insights into the Economic Toll of Cell Line Misidentification: A Comprehensive Review (2026) · cites this paper as [54]

    Section 1.2 says dPCR partitions a sample so that each partition contains few or no target sequences; it does not say the sample must contain little DNA. Section 3.2 describes detection at high molecule concentrations and says the number of target sequences detected can exceed the number of partitions. Low DNA quantity is therefore not a requirement stated by this source. (LLM, conf=0.82)

    Checked by a reasoning model — may be wrong.

    Experimental — accuracy not yet measured

  • dPCR provides exquisite sensitivity for absolute quantification of target nucleic acids.

    “It requires low DNA quantities and provides exquisite sensitivity for absolute quantification of target nucleic acids [54,55,56].”

    From Genetic Insights into the Economic Toll of Cell Line Misidentification: A Comprehensive Review (2026) · cites this paper as [54]

    Section 1.2 states that dPCR enables absolute quantification of target nucleic acids. But Section 7 says dPCR has lower sensitivity for absolute quantification than qPCR, so the review does not support the unqualified description of its sensitivity as “exquisite.” (LLM, conf=0.88)

    Experimental — accuracy not yet measured

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Cited by

Sorted by citations

Authors
Quan, P.-L., Sauzade, M., & Brouzes, E.
Affiliations
Stony Brook University (US)
Corresponding author
Brouzes, Eric
ORCID
Venue
Sensors
Publisher
MDPI AG
Published
2018-04-20
Volume
18
Issue
4
Pages
1271
ISSN
1424-8220 (ISSN-L)
DOI
10.3390/s18041271
PMC
PMC5948698
OpenAlex
W2799273553
Cited by
664
References
12
Citations by year
2026: 74 · 2025: 91 · 2024: 95 · 2023: 73 · 2022: 93 · 2021: 94 · 2020: 92 · 2019: 41 · 2018: 10
FWCI
5.79
Type
journal article
Language
en
Open access
Licence
Locations
  • Sensors (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
NHI- NCI (RO1 CA181595)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Molecular Biology Techniques and Applications
  • Physical Sciences › Engineering › Biomedical Engineering › Microfluidic and Capillary Electrophoresis Applications
  • Physical Sciences › Engineering › Biomedical Engineering › Innovative Microfluidic and Catalytic Techniques Innovation
Keywords
digital PCR, absolute quantification, Poisson distribution, multiplexing, isothermal amplification, nucleic acid quantification

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