ArticleAnalytical and bioanalytical chemistry2017
Droplet volume variability as a critical factor for accuracy of absolute quantification using droplet digital PCR.
Article in Analytical and bioanalytical chemistry, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 35 papers.
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Who cites it
35 citing papers in PubMed, 94 citations in OpenAlex.
- Development of reference material from Rift Valley fever virus RNA.Virology journal · 2026Article
- Droplet Digital CRISPR for Nucleic Acid Detection.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Review
- Comparison of two digital PCR platforms for quantification of genetically modified soybean events.GM crops & food · 2025Article
- A method to validate viral copy-number assay involving a hybrid amplicon and duplex droplet digital PCR.Molecular therapy. Methods & clinical development · 2025Article
- Absolute quantification of rare gene targets in limited samples using crude lysate and ddPCR.Scientific reports · 2025Article
- Considerations for the Successful Detection and Quantification of Genetically Modified Events in Grain and Food Samples Using Multiplex Digital PCR.Foods (Basel, Switzerland) · 2024Article
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- Fast and Accurate Multiplex Identification and Quantification of Seven Genetically Modified Soybean Lines Using Six-Color Digital PCR.Foods (Basel, Switzerland) · 2023Article
- DNA-Binding Magnetic Nanoreactor Beads for Digital PCR Analysis.Analytical chemistry · 2023Article
- Large-scale F0 CRISPR screens in vivo using MIC-Drop.Nature protocols · 2023Review
- Evolution of tuberculosis diagnostics: From molecular strategies to nanodiagnostics.Tuberculosis (Edinburgh, Scotland) · 2023Review
- Evaluation of DNA Extraction Methods for Reliable Quantification ofBiosensors · 2023Article
- Body Map of Droplet Distributions During Oropharyngeal Suction to Protect Health Care Workers From Airborne Diseases.Journal of perianesthesia nursing : official journal of the American Society of PeriAnesthesia Nurses · 2023Review
- Cost-Effective Droplet Generator for Portable Bio-Applications.Micromachines · 2023Article
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- Quantification of cell-free circulating mitochondrial DNA copy number variation in hepatocellular carcinoma.Revista da Associacao Medica Brasileira (1992) · 2022Article
- Assessing the Variability of Cell-Associated HIV DNA Quantification through a Multicenter Collaborative Study.Microbiology spectrum · 2022Article
- Screening strategies and laboratory assays to support Plasmodium falciparum histidine-rich protein deletion surveillance: where we are and what is needed.Malaria journal · 2022Review
Corrections and comments
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Authors and funding
9 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Accurate and precise nucleic-acid quantification is crucial for clinical and diagnostic decisions, as overestimation or underestimation can lead to misguided treatment of a disease or incorrect labelling of the products. Digital PCR is one of the best tools for absolute nucleic-acid copy-number determination. However, digital PCR needs to be well characterised in terms of accuracy and sources of uncertainty. With droplet digital PCR, discrepancies between the droplet volume assigned by the manufacturer and measured by independent laboratories have already been shown in previous studies. In the present study, we report on the results of an inter-laboratory comparison of different methods for droplet volume determination that is based on optical microscopy imaging and is traceable to the International System of Units. This comparison was conducted on the same DNA material, with the examination of the influence of parameters such as droplet generators, supermixes, operators, inter-cartridge and intra-cartridge variability, and droplet measuring protocol. The mean droplet volume was measured using a QX200™ AutoDG™ Droplet Digital™ PCR system and two QX100™ Droplet Digital™ PCR systems. The data show significant volume differences between these two systems, as well as significant differences in volume when different supermixes are used. We also show that both of these droplet generator systems produce droplets with significantly lower droplet volumes (13.1%, 15.9%, respectively) than stated by the manufacturer and previously measured by other laboratories. This indicates that to ensure precise quantification, the droplet volumes should be assessed for each system.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.