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Sensors 2018, 18(4), 1271; https://doi.org/10.3390/s18041271

dPCR: A Technology Review

1
Department of Biomedical Engineering, Stony Brook University, Stony Brook, NY 11794, USA
2
Laufer Center for Physical and Quantitative Biology, Stony Brook University, Stony Brook, NY 11794, USA
These authors contributed equally to this work.
*
Author to whom correspondence should be addressed.
Received: 10 March 2018 / Revised: 13 April 2018 / Accepted: 15 April 2018 / Published: 20 April 2018
(This article belongs to the Special Issue Bio-MEMS for Precision Medicine)
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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. View Full-Text
Keywords: absolute quantification; arrays of microwells; digital PCR; dPCR; droplet microfluidics; microfluidics; microfluidic chambers; microfluidic technologies; on-chip valves; partitioning; quantitative real-time PCR; qPCR; real-time PCR absolute quantification; arrays of microwells; digital PCR; dPCR; droplet microfluidics; microfluidics; microfluidic chambers; microfluidic technologies; on-chip valves; partitioning; quantitative real-time PCR; qPCR; real-time PCR
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).
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Quan, P.-L.; Sauzade, M.; Brouzes, E. dPCR: A Technology Review. Sensors 2018, 18, 1271.

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