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dPCR: A Technology Review

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 microrea...

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Autores principales: Quan, Phenix-Lan, Sauzade, Martin, Brouzes, Eric
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5948698/
https://www.ncbi.nlm.nih.gov/pubmed/29677144
http://dx.doi.org/10.3390/s18041271
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author Quan, Phenix-Lan
Sauzade, Martin
Brouzes, Eric
author_facet Quan, Phenix-Lan
Sauzade, Martin
Brouzes, Eric
author_sort Quan, Phenix-Lan
collection PubMed
description 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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spelling pubmed-59486982018-05-17 dPCR: A Technology Review Quan, Phenix-Lan Sauzade, Martin Brouzes, Eric Sensors (Basel) Review 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. MDPI 2018-04-20 /pmc/articles/PMC5948698/ /pubmed/29677144 http://dx.doi.org/10.3390/s18041271 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Review
Quan, Phenix-Lan
Sauzade, Martin
Brouzes, Eric
dPCR: A Technology Review
title dPCR: A Technology Review
title_full dPCR: A Technology Review
title_fullStr dPCR: A Technology Review
title_full_unstemmed dPCR: A Technology Review
title_short dPCR: A Technology Review
title_sort dpcr: a technology review
topic Review
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5948698/
https://www.ncbi.nlm.nih.gov/pubmed/29677144
http://dx.doi.org/10.3390/s18041271
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