ArticleScientific reports2019
Optimisation of robust singleplex and multiplex droplet digital PCR assays for high confidence mutation detection in circulating tumour DNA.
Article in Scientific reports, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 52 papers.
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Who cites it
52 citing papers in PubMed.
- An ultrasensitive aptamer-based fluorescent biosensor for luteinizing hormone with mutually orthogonal DNAzyme and self-replication CHA amplification.Analytical and bioanalytical chemistry · 2026Article
- Optimized Viability-ddPCR with Triton X-100 Enhancement for Selective Detection of LiveJournal of microbiology and biotechnology · 2026Article
- Robust dPCR Hyper-Plexing Approach by Color Combination for Detection of 17 ESR1 Mutations in Breast Cancer Patient Samples.Methods in molecular biology (Clifton, N.J.) · 2026Article
- The Clinical Utility of Cell-Free DNA in Brain Tumor Management: A Comprehensive Review.Journal of molecular neuroscience : MN · 2025Review
- Minimal Residual Disease Detection: Bridging Molecular and Clinical Strategies for Recurrence Prevention in Gynecologic Cancers.International journal of molecular sciences · 2025Review
- Comparative Analysis of Two Digital PCR Platforms for Detecting DNA Methylation in Patient Samples.Cell biochemistry and function · 2025Article
- Prognostic Significance of the Comprehensive Biomarker Analysis in Colorectal Cancer.Life (Basel, Switzerland) · 2025Review
- Pre-Amplification of Cell-Free DNA: Balancing Amplification Errors with Enhanced Sensitivity.Biomolecules · 2025Article
- Mosaic X-linked adrenoleukodystrophy in males identified by newborn screening and next-generation sequencing.NPJ genomic medicine · 2025Article
- Circulating tumor DNA in colorectal cancer: biology, methods and applications.Discover oncology · 2025Review
- Absolute quantification of rare gene targets in limited samples using crude lysate and ddPCR.Scientific reports · 2025Article
- Advancing artemisinin resistance monitoring using a high sensitivity ddPCR assay for Pfkelch13 mutation detection in Asia.Scientific reports · 2025Article
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- Detection of EGFR mutations in patients with suspected lung cancer using paired tissue-plasma testing: a prospective comparative study with plasma ddPCR assay.Scientific reports · 2024Article
- Generic Reporter Sets for Colorimetric Multiplex dPCR Demonstrated with 6-Plex SNP Quantification Panels.International journal of molecular sciences · 2024Article
- Detection of the JAK2 V617F Mutation in Urinary Cell-free DNA in Patients with Myeloproliferative Neoplasms.Internal medicine (Tokyo, Japan) · 2024Article
- Droplet digital PCR analysis of CDH13 methylation status in Slovak women with invasive ductal breast cancer.Scientific reports · 2024Article
- Optimization and application of digital droplet PCR for the detection of SARS-CoV-2 in saliva specimen using commercially available kit.Biology methods & protocols · 2024Article
- Circulating Tumour Cells: Detection and Application in Advanced Non-Small Cell Lung Cancer.International journal of molecular sciences · 2023Review
Corrections and comments
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Liquid biopsies offer the potential to monitor cancer response and resistance to therapeutics in near real-time. However, the plasma cell free DNA (cfDNA) level can be low and the fraction of circulating tumour DNA (ctDNA) bearing a mutation - lower still. Detection of tumour-derived mutations in ctDNA is thus challenging and requires highly sensitive and specific assays. Droplet digital PCR (ddPCR) is a technique that enables exquisitely sensitive detection and quantification of DNA/RNA markers from very limiting clinical samples, including plasma. The Bio-Rad QX200 ddPCR system provides absolute quantitation of target DNA molecules using fluorescent dual-labelled probes. Critical to accurate sample analysis are validated assays that are highly specific, reproducible, and with known performance characteristics, especially with respect to false positives. We present a systematic approach to the development and optimisation of singleplex and multiplex ddPCR assays for the detection of point mutations with a focus on ensuring extremely low false positives whilst retaining high sensitivity. We also present a refined method to determine cfDNA extraction efficiency allowing for more accurate extrapolation of mutational levels in source samples. We have applied these approaches to successfully analyse many ctDNA samples from multiple clinical studies and generated exploratory data of high quality.
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