ArticleThe Journal of molecular diagnostics : JMD2011
Optimized allele-specific real-time PCR assays for the detection of common mutations in KRAS and BRAF.
Article in The Journal of molecular diagnostics : JMD, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 44 papers, 3 of them syntheses 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
44 citing papers in PubMed, 3 syntheses or guidelines pooled it.
- Molecular Biomarkers for the Evaluation of Colorectal Cancer: Guideline From the American Society for Clinical Pathology, College of American Pathologists, Association for Molecular Pathology, and American Society of Clinical Oncology.The Journal of molecular diagnostics : JMD · 2017Guideline
- Molecular Biomarkers for the Evaluation of Colorectal Cancer.American journal of clinical pathology · 2017Guideline
- Pooled it
- A quantitative polymerase chain reaction approach for distinguishing the geographical origin ofFood science and biotechnology · 2025Article
- Unlocking the Genetic Secrets of Pancreatic Cancer:Cancers · 2025Review
- Liquid biopsies in cancer.Molecular biomedicine · 2025Review
- Rapid intraoperative amplicon sequencing of CNS tumor markers.Computational and structural biotechnology journal · 2024Article
- Inhibition of ADAM9 promotes the selective degradation of KRAS and sensitizes pancreatic cancers to chemotherapy.Nature cancer · 2024Article
- Phosphoramidate Azole Oligonucleotides for Single Nucleotide Polymorphism Detection by PCR.International journal of molecular sciences · 2024Article
- Efficacy of bupropion and varenicline genetic markers in choosing pharmacological treatment for smoking cessation, and implications for combining drugs: A randomized controlled trial - GENTSMOKING.Tobacco induced diseases · 2024Article
- Screening and validation of double allele-specific binding F-primers for the measurement of antihypertensive pharmacogenomics.Frontiers in medicine · 2023Article
- Liquid biopsy at the frontier of detection, prognosis and progression monitoring in colorectal cancer.Molecular cancer · 2022Review
- T7 Endonuclease I-mediated voltammetric detection of KRAS mutation coupled with horseradish peroxidase for signal amplification.Mikrochimica acta · 2022Article
- The Opportunities and Challenges of Molecular Tagging Next-Generation Sequencing in Liquid Biopsy.Molecular diagnosis & therapy · 2021Article
- Single-copy detection of somatic variants from solid and liquid biopsy.Scientific reports · 2021Article
- Allele-Specific PCR forDiagnostics (Basel, Switzerland) · 2020Article
- A highly sensitive and specific real-time quantitative PCR for BRAF V600E/K mutation screening.Scientific reports · 2020Article
- Article
- Polymerase-amplified release of ATP (POLARA) for detecting single nucleotide variants in RNA and DNA.Chemical science · 2019Article
- Identification of Thermus aquaticus DNA polymerase variants with increased mismatch discrimination and reverse transcriptase activity from a smart enzyme mutant library.Scientific reports · 2019Article
Corrections and comments
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Authors and funding
7 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Mutations in the oncogenes KRAS and BRAF have been identified as prognostic factors in patients with colorectal diseases and as predictors of negative outcome in epidermal growth factor receptor-targeted therapies. Therefore, accurate mutation detection in both genes, KRAS and BRAF, is of increasing clinical relevance. We aimed at optimizing allele-specific real-time PCR assays for the detection of common mutations in KRAS and the BRAF Val600Glu mutation using allele-specific PCR primers for allelic discrimination and probes (TaqMan) for quantification. Each reaction mix contains a co-amplified internal control to exclude false-negative results. Allele-specific real-time PCR assays were evaluated on plasmid model systems providing a mutation detection limit of 10 copies of mutant DNA in proportions as low as 1% of the total DNA. Furthermore, we analyzed 125 DNA samples prepared from archived, formalin-fixed, paraffin-embedded colorectal carcinomas and compared results with those obtained from direct-sequence analysis. All mutations determined by sequence analysis could be recovered by allele-specific PCR assays. In addition, allele-specific PCR assays clearly identified three additional samples affected by a mutation. We propose these allele-specific real-time PCR assays as a low-cost and fast diagnostic tool for accurate detection of KRAS and BRAF mutations that can be applied to clinical samples.
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