ArticleVirchows Archiv : an international journal of pathology2012
Analytical performance of a PCR assay for the detection of KRAS mutations (codons 12/13 and 61) in formalin-fixed paraffin-embedded tissue samples of colorectal carcinoma.
Article in Virchows Archiv : an international journal of pathology, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 27 papers.
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27 citing papers in PubMed, 52 citations in OpenAlex.
- Erlotinib is effective in pancreatic cancer with epidermal growth factor receptor mutations: a randomized, open-label, prospective trial.Oncotarget · 2015Trial
- 'The Reports of My Death Are Greatly Exaggerated'-Evaluating the Effect of Necrosis onCancers · 2024Article
- Concomitant KRAS mutations attenuate sensitivity of non-small cell lung cancer cells to KRAS G12C inhibition.Scientific reports · 2022Article
- KRAS mutations in patients with colorectal cancer in Libya.Molecular and clinical oncology · 2021Article
- PNA Clamping in Nucleic Acid Amplification Protocols to Detect Single Nucleotide Mutations Related to Cancer.Molecules (Basel, Switzerland) · 2020Review
- Clinical Evaluation of IntelliPlex™ KRAS G12/13 Mutation Kit for Detection of KRAS Mutations in Codon 12 and 13: A Novel Multiplex Approach.Molecular diagnosis & therapy · 2019Article
- Rapid and accurate detection ofOncotarget · 2017Article
- Current companion diagnostics in advanced colorectal cancer; getting a bigger and better piece of the pie.Journal of gastrointestinal oncology · 2017Review
- Key differences between 13 KRAS mutation detection technologies and their relevance for clinical practice.ESMO open · 2017Article
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- Novel Approach for Clinical Validation of the cobas KRAS Mutation Test in Advanced Colorectal Cancer.Molecular diagnosis & therapy · 2016Article
- Diagnostic RAS mutation analysis by polymerase chain reaction (PCR).Biomolecular detection and quantification · 2016Article
- Article
- BRAF, PIK3CA, and HER2 Oncogenic Alterations According to KRAS Mutation Status in Advanced Colorectal Cancers with Distant Metastasis.PloS one · 2016Article
- Article
- Detection of KRAS, NRAS and BRAF by mass spectrometry - a sensitive, reliable, fast and cost-effective technique.Diagnostic pathology · 2015Article
- Eprobe-mediated screening system for somatic mutations in the KRAS locus.Oncology reports · 2015Article
- c-MYC Copy-Number Gain Is an Independent Prognostic Factor in Patients with Colorectal Cancer.PloS one · 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
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7 authors at 1 institution in 1 country.
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No grant is acknowledged in the PubMed record.
Abstract
KRAS mutation testing is mandatory before prescribing anti-epidermal growth factor monoclonal antibodies in the treatment of advanced colorectal cancer. We describe the performance of a TaqMelt polymerase chain reaction (PCR) assay-the cobas® KRAS Mutation Test-designed to detect 19 mutations in codons 12, 13, and 61. The limit of detection was determined using DNA blends from cell lines, plasmids, and formalin-fixed paraffin-embedded tissue specimens. Assay performance was compared to Sanger sequencing using a panel of 188 specimens. Discordant specimens were subjected to next generation pyrosequencing (454). Assay repeatability was assessed using a panel of six specimens. A >95% correct mutation call rate was obtained in all specimen types with ~5% mutant alleles at DNA inputs of 0.8-6.3 ng per PCR reaction; 100% detection rate was observed at the recommended DNA input of 50 ng. The positive percent agreement with Sanger was 97.5% (79/81) for codons 12/13 and 85.7% (6/7) for codon 61. Negative percent agreement was 94.4% (101/107) for codon 12/13 and 99.4% (180/181) for codon 61. Nine of 10 discordant specimens yielded 454 results consistent with the cobas® results. With repeated testing, the assay showed a correct call rate of 100% (192/192) for all operators, instruments, reagent lots, and days tested. The cobas® test detects KRAS mutations in codons 12, 13, and 61 at a limit of detection of <5%. The PCR assay was more sensitive and specific than Sanger sequencing, and performance was highly reproducible. Test performance was not influenced by various endogenous interfering substances or common gut microbes.
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