ArticleScientific reports2023
Efficient and accurate KRAS genotyping using digital PCR combined with melting curve analysis for ctDNA from pancreatic cancer patients.
Article in Scientific reports, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed, 21 citations in OpenAlex.
- KRAS Mutations in Duodenal Lavage Fluid After Secretin Stimulation for Detection of Pancreatic Cancer.Annals of surgery · 2026Article
- High-Sensitivity ctDNA Analysis Uncovers Relevant Signals Missed by NGS in Pancreatic Cancer.Clinical cancer research : an official journal of the American Association for Cancer Research · 2026Article
- Quantitative KRAS mutational burden by next-generation sequencing predicts outcomes in localized pancreatic cancer.Surgery · 2026Article
- Evidence Mapping of ctDNA Reporting in Pancreatic Ductal Adenocarcinoma: Toward a Shared Quantitative Language for ctDNA.Cancers · 2026Review
- Review
- Highly multiplexed digital PCR assay for simultaneous quantification of variant allele frequencies and copy number alterations of KRAS and GNAS in pancreatic cancer precursors.Molecular oncology · 2025Article
- [Advances in Diagnosis and Targeted Therapy of KRASG12C Mutant Non-small Cell Lung Cancer].Zhongguo fei ai za zhi = Chinese journal of lung cancer · 2025Review
- Analytical techniques for nucleic acid and protein detection with single-molecule sensitivity.Experimental & molecular medicine · 2025Review
- Utility of circulating tumor DNA in secondary liver malignancies: What we know and what is to come.Journal of surgical oncology · 2025Review
- Coupling enzymatic digestion with nanopore sensing for low-abundance EGFR-L858R mutation detection.Frontiers in bioengineering and biotechnology · 2025Article
- High somatic mutations in circulating tumor DNA predict response of metastatic pancreatic ductal adenocarcinoma to first-line nab-paclitaxel plus S-1: prospective study.Journal of translational medicine · 2024Article
- Potential of Alkalization Therapy for the Management of Metastatic Pancreatic Cancer: A Retrospective Study.Cancers · 2023Article
- Pancreatic cancer surgery.BMC surgery · 2023Article
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Authors and funding
14 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
A highly sensitive and highly multiplexed quantification technique for nucleic acids is necessary to predict and evaluate cancer treatment by liquid biopsy. Digital PCR (dPCR) is a highly sensitive quantification technique, but conventional dPCR discriminates multiple targets by the color of the fluorescent dye of the probe, which limits multiplexing beyond the number of colors of fluorescent dyes. We previously developed a highly multiplexed dPCR technique combined with melting curve analysis. Herein, we improved the detection efficiency and accuracy of multiplexed dPCR with melting curve analysis to detect KRAS mutations in circulating tumor DNA (ctDNA) prepared from clinical samples. The mutation detection efficiency was increased from 25.9% of the input DNA to 45.2% by shortening the amplicon size. The limit of detection of mutation was improved from 0.41 to 0.06% by changing the mutation type determination algorithm for G12A, resulting in a limit of detection of less than 0.2% for all the target mutations. Then, ctDNA in plasma from pancreatic cancer patients was measured and genotyped. The measured mutation frequencies correlated well with those measured by conventional dPCR, which can measure only the total frequency of KRAS mutants. KRAS mutations were detected in 82.3% of patients with liver or lung metastasis, which was consistent with other reports. Accordingly, this study demonstrated the clinical utility of multiplex dPCR with melting curve analysis to detect and genotype ctDNA from plasma with sufficient sensitivity.
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