ArticleThe Journal of molecular diagnostics : JMD2010
Detection of low-level KRAS mutations using PNA-mediated asymmetric PCR clamping and melting curve analysis with unlabeled probes.
Article in The Journal of molecular diagnostics : JMD, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 25 papers, 1 of them a synthesis that pooled it.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
25 citing papers in PubMed, 1 synthesis or guideline pooled it, 55 citations in OpenAlex.
- Pooled it
- Sequential CRISPR-EspCas9-Mediated Wild-Type Depletion Enhances the Detection Sensitivity of Rare Mutations for Canine Liquid Biopsy Application.Biosensors · 2026Article
- Identification of breast cancer-associated PIK3CA H1047R mutation in blood circulation using an asymmetric PCR assay.PloS one · 2024Article
- Sequence-Based Platforms for Discovering Biomarkers in Liquid Biopsy of Non-Small-Cell Lung Cancer.Cancers · 2023Review
- Clinical Application Value of Circulating Cell-free DNA in Hepatocellular Carcinoma.Frontiers in molecular biosciences · 2021Review
- Harnessing cell-free DNA: plasma circulating tumour DNA for liquid biopsy in genitourinary cancers.Nature reviews. Urology · 2020Review
- SLAM-MS: Mutation scanning of stem-loop amplicons with TaqMan probes by quantitative DNA melting analysis.Scientific reports · 2020Article
- Role of oncogenic KRAS in the diagnosis, prognosis and treatment of pancreatic cancer.Nature reviews. Gastroenterology & hepatology · 2020Review
- PNA Clamping in Nucleic Acid Amplification Protocols to Detect Single Nucleotide Mutations Related to Cancer.Molecules (Basel, Switzerland) · 2020Review
- Genomic Analysis Identifies Mutations Concerning Drug-Resistance and Beijing Genotype in Multidrug-ResistantFrontiers in microbiology · 2020Article
- Article
- [Progress in Non-invasive Detection of EGFR Mutation in Non-small Cell Lung Cancer].Zhongguo fei ai za zhi = Chinese journal of lung cancer · 2018Review
- Integrating liquid biopsies into the management of cancer.Nature reviews. Clinical oncology · 2017Review
- Molecular Testing for Gastrointestinal Cancer.Journal of pathology and translational medicine · 2017Review
- Eprobe-mediated screening system for somatic mutations in the KRAS locus.Oncology reports · 2015Article
- Single-Tubed Wild-Type Blocking Quantitative PCR Detection Assay for the Sensitive Detection of Codon 12 and 13 KRAS Mutations.PloS one · 2015Article
- Article
- KRAS mutations in tumor tissue and plasma by different assays predict survival of patients with metastatic colorectal cancer.Journal of experimental & clinical cancer research : CR · 2014Article
- Stress-induced DNA damage: a case study in diffuse large B-cell lymphoma.Journal of the Royal Society, Interface · 2014Article
- Real-time bidirectional pyrophosphorolysis-activated polymerization for quantitative detection of somatic mutations.PloS one · 2014Article
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
7 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Detection of somatic mutations in clinical cancer specimens is often hampered by excess wild-type DNA. The aim of this study was to develop a simple and economical protocol without using fluorescent probes to detect low-level mutations. In this study, we combined peptide nucleic acid (PNA)-clamping PCR with asymmetric primers and a melting curve analysis using an unlabeled detection probe. PNA-clamping PCR, which suppressed amplification of the wild-type allele, was more sensitive for KRAS codon 12 mutation detection than nonclamping PCR in 5 different mutant cell lines. Three detection probes were tested (a perfectly matched antisense, a mismatched antisense, and a mismatched sense), and the mismatched sense detection probe showed the highest sensitivity (0.1% mutant detection) under clamping conditions. With this probe, we were able to detect not only the perfectly matched KRAS mutation, but also 4 other mismatched mutations of KRAS. We then applied this protocol to 10 human colon cancer tissues with KRAS codon 12 mutations, successfully detecting the mutations in all of them. Our data indicate that the combination of perfectly matched antisense PNA and a mismatched sense detection probe can detect KRAS mutations with a high sensitivity in both cell lines and human tissues. Moreover, this study might prove an easily applicable protocol for the detection of low-level mutations in other cancer genes.
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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.