ArticleNucleic acids research2007
FLAG assay as a novel method for real-time signal generation during PCR: application to detection and genotyping of KRAS codon 12 mutations.
Article in Nucleic acids research, 2007. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 25 papers.
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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, 50 citations in OpenAlex.
- Pyrimidine-Dependent UV-Mediated Cross-Linking Magnifies Minor Genetic or Epigenetic Changes in Clinical Samples.Clinical chemistry · 2024Article
- Enzymatic Methods for Mutation Detection in Cancer Samples and Liquid Biopsies.International journal of molecular sciences · 2023Review
- Review
- Mutation enrichment in human DNA samples via UV-mediated cross-linking.Nucleic acids research · 2022Article
- Nuclease-Assisted, Multiplexed Minor-Allele Enrichment: Application in Liquid Biopsy of Cancer.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Untangling the KRAS mutated lung cancer subsets and its therapeutic implications.Molecular biomedicine · 2021Review
- Denaturation-Enhanced Droplet Digital PCR for Liquid Biopsies.Clinical chemistry · 2018Article
- A novel RFLP-ARMS TaqMan PCR-based method for detecting the BRAF V600E mutation in melanoma.Oncology letters · 2018Article
- Multiplexed Elimination of Wild-Type DNA and High-Resolution Melting Prior to Targeted Resequencing of Liquid Biopsies.Clinical chemistry · 2017Article
- Methylation-sensitive enrichment of minor DNA alleles using a double-strand DNA-specific nuclease.Nucleic acids research · 2017Article
- Elimination of unaltered DNA in mixed clinical samples via nuclease-assisted minor-allele enrichment.Nucleic acids research · 2016Article
- RAS testing in metastatic colorectal cancer: advances in Europe.Virchows Archiv : an international journal of pathology · 2016Review
- Article
- Article
- Enhanced ratio of signals enables digital mutation scanning for rare allele detection.The Journal of molecular diagnostics : JMD · 2015Article
- Article
- A novel fully automated molecular diagnostic system (AMDS) for colorectal cancer mutation detection.PloS one · 2013Article
- A multiplex SNaPshot assay as a rapid method for detecting KRAS and BRAF mutations in advanced colorectal cancers.The Journal of molecular diagnostics : JMD · 2011Article
- Biotinylated probe isolation of targeted gene region improves detection of T790M epidermal growth factor receptor mutation via peptide nucleic acid-enriched real-time PCR.Clinical chemistry · 2011Article
- Sensitive and specific KRAS somatic mutation analysis on whole-genome amplified DNA from archival tissues.The Journal of molecular diagnostics : JMD · 2010Article
Corrections and comments
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Authors and funding
4 authors at 2 institutions in 1 country.
Funding
Abstract
Real-time signal generation methods for detection and characterization of low-abundance mutations in genomic DNA are powerful tools for cancer diagnosis and prognosis. Mutations in codon 12 of the oncogene KRAS, for example, are frequently found in several types of human cancers. We have developed a novel real-time PCR technology, FLAG (FLuorescent Amplicon Generation) and adapted it for simultaneously (i) amplifying mutated codon 12 KRAS sequences, (ii) monitoring in real-time the amplification and (iii) genotyping the exact nucleotide alteration. FLAG utilizes the exceptionally thermostable endonuclease PspGI for real-time signal generation by cleavage of quenched fluorophores from the 5'-end of the PCR products and, concurrently, for selecting KRAS mutations over wild type. By including peptide-nucleic-acid probes in the reaction, simultaneous genotyping is achieved that circumvents the requirement for sequencing. FLAG enables high-throughput, closed-tube KRAS mutation detection down to approximately 0.1% mutant-to-wild type. The assay was validated on model systems and compared with allele-specific PCR sequencing for screening 27 cancer specimens. Diverse applications of FLAG for real-time PCR or genotyping applications in cancer, virology or infectious diseases are envisioned.
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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.