Absolute quantification by droplet digital PCR versus analog real-time PCR
Open accessAbstract
Nanoliter-sized droplet technology paired with digital PCR (ddPCR) holds promise for highly precise, absolute nucleic acid quantification. Our comparison of microRNA quantification by ddPCR and real-time PCR revealed greater precision (coefficients of variation decreased 37-86%) and improved day-to-day reproducibility (by a factor of seven) of ddPCR but with comparable sensitivity. When we applied ddPCR to serum microRNA biomarker analysis, this translated to superior diagnostic performance for identifying individuals with cancer.
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Recent studies comparing qPCR and dPCR reported improved precision and reproducibility with dPCR.
“Recent studies have compared the two technologies and reported improved precision and reproducibility with dPCR (11).”
From Analysis of Extracellular RNA by Digital PCR (2014) · cites this paper as [11]
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“Droplet Digital PCR (ddPCR) is a recent technology that has become commercially available since 2011 9, 10 .”
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“Recent studies have compared the two technologies and reported improved precision and reproducibility with dPCR (11).”
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- Authors
- Hindson, C. M., Chevillet, J. R., Briggs, H. A., Gallichotte, E. N., Ruf, I. K., Hindson, B. J., Vessella, R. L., & Tewari, M.
- Affiliations
- Bio-Rad (United States) (US)
- Fred Hutch Cancer Center (US)
- University of Washington (US)
- Corresponding author
- Tewari, Muneesh
- Venue
- Nature Methods
- Published
- 2013-09-01
- Volume
- 10
- Issue
- 10
- Pages
- 1003-1005
- ISSN
- 1548-7091 (ISSN-L)
- 1548-7105
- PMC
- PMC4118677
- OpenAlex
- W2042194227
- Cited by
- 1,488
- References
- 2
- Citations by year
- 2026: 83 · 2025: 120 · 2024: 131 · 2023: 133 · 2022: 128 · 2021: 164 · 2020: 144 · 2019: 112 · 2018: 123 · 2017: 116 · 2016: 116 · 2015: 68 · 2014: 43 · 2013: 3
- FWCI
- 34.37
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature Methods (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institutes of Health (5p50ca083636-15)
- National Cancer Institute (P50-CA083636, P50 CA83636, P01CA085859, CA100606, U01CA157703, P50CA097186, P01CA85859)
- National Institute of Diabetes and Digestive and Kidney Diseases (R01DK085714)
- National Institute of Biomedical Imaging and Bioengineering (R01EB010106)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Molecular Biology Techniques and Applications
- Physical Sciences › Engineering › Biomedical Engineering › Innovative Microfluidic and Catalytic Techniques Innovation
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Advanced biosensing and bioanalysis techniques
- Keywords
- droplet digital PCR, absolute quantification, real-time PCR comparison, microRNA quantification, nanoliter droplets, precision and reproducibility, detection sensitivity, serum microRNA biomarker, cancer diagnosis
- MeSH terms
- Humans, Male, Prostatic Neoplasms, Sensitivity and Specificity, Biomarkers, Tumor, Reproducibility of Results, MicroRNAs, Limit of Detection, Real-Time Polymerase Chain Reaction
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