ArticlePloS one2013
Methods for applying accurate digital PCR analysis on low copy DNA samples.
Article in PloS one, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 83 papers.
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Who cites it
83 citing papers in PubMed, 155 citations in OpenAlex.
- Portable qPCR and ddPCR Diagnostics for Cercospora Leaf Spot: Integrating Field and Laboratory Detection ofJournal of fungi (Basel, Switzerland) · 2026Article
- Digital PCR in forensic genetics: principles, applications, and challenges.International journal of legal medicine · 2026Review
- Lessons learned from applying eDNA surveying to diadromous fish detection across the Northeast Atlantic region.Journal of fish biology · 2026Article
- Implementation and Validation of a Limiting Component Quantification Method for qPCR.International journal of molecular sciences · 2026Article
- A method to validate viral copy-number assay involving a hybrid amplicon and duplex droplet digital PCR.Molecular therapy. Methods & clinical development · 2025Article
- Establishment and application of a dual chip digital PCR assay for detection of PDCoV and PEDV.Frontiers in veterinary science · 2025Article
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- Advancing Pathogen Identification: The Role of Digital PCR in Enhancing Diagnostic Power in Different Settings.Diagnostics (Basel, Switzerland) · 2024Review
- A multiplex digital PCR assay for detection and quantitation of porcine circovirus type 2 and type 3.Archives of virology · 2024Article
- Quantification of low-level human cytomegalovirus and Epstein-Barr virus DNAemia by digital PCR.Journal of virological methods · 2024Article
- Research on quantitative detection technology of raccoon-derived ingredient adulteration in sausage products.Food science & nutrition · 2024Article
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- Triplex Crystal Digital PCR for the Detection and Differentiation of the Wild-Type Strain and the MGF505-2R and I177L Gene-Deleted Strain of African Swine Fever Virus.Pathogens (Basel, Switzerland) · 2023Article
- Development of a Multiplex Crystal Digital RT-PCR for Differential Detection of Classical, Highly Pathogenic, and NADC30-like Porcine Reproductive and Respiratory Syndrome Virus.Animals : an open access journal from MDPI · 2023Article
- Development of the Droplet Digital PCR Method for the Detection and Quantification of Erwinia pyrifoliae.The plant pathology journal · 2023Article
- Optimizing Molecular Minimal Residual Disease Analysis in Adult Acute Lymphoblastic Leukemia.Cancers · 2023Review
- Research Topic: Measurable Residual Disease in Hematologic Malignancies. Can digital droplet PCR improve measurable residual disease monitoring in chronic lymphoid malignancies?Frontiers in oncology · 2023Review
- Single-Cell Analysis of circRNA Using ddPCR.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Assessing the Variability of Cell-Associated HIV DNA Quantification through a Multicenter Collaborative Study.Microbiology spectrum · 2022Article
23 more citing papers are in PubMed but not listed here.
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Digital PCR (dPCR) is a highly accurate molecular approach, capable of precise measurements, offering a number of unique opportunities. However, in its current format dPCR can be limited by the amount of sample that can be analysed and consequently additional considerations such as performing multiplex reactions or pre-amplification can be considered. This study investigated the impact of duplexing and pre-amplification on dPCR analysis by using three different assays targeting a model template (a portion of the Arabidopsis thaliana alcohol dehydrogenase gene). We also investigated the impact of different template types (linearised plasmid clone and more complex genomic DNA) on measurement precision using dPCR. We were able to demonstrate that duplex dPCR can provide a more precise measurement than uniplex dPCR, while applying pre-amplification or varying template type can significantly decrease the precision of dPCR. Furthermore, we also demonstrate that the pre-amplification step can introduce measurement bias that is not consistent between experiments for a sample or assay and so could not be compensated for during the analysis of this data set. We also describe a model for estimating the prevalence of molecular dropout and identify this as a source of dPCR imprecision. Our data have demonstrated that the precision afforded by dPCR at low sample concentration can exceed that of the same template post pre-amplification thereby negating the need for this additional step. Our findings also highlight the technical differences between different templates types containing the same sequence that must be considered if plasmid DNA is to be used to assess or control for more complex templates like genomic DNA.
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Registered trials
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.