ArticleThe Journal of molecular diagnostics : JMD2008
High-fidelity DNA polymerase enhances the sensitivity of a peptide nucleic acid clamp PCR assay for K-ras mutations.
Article in The Journal of molecular diagnostics : JMD, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
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Who cites it
18 citing papers in PubMed, 43 citations in OpenAlex.
- Clinical relevance of circulating KRAS mutated DNA in plasma from patients with advanced pancreatic cancer.Molecular oncology · 2016Trial
- Toward Safer Biotherapeutics: Expression and Characterization of a Humanized Chimeric L-Asparaginase inInternational journal of molecular sciences · 2025Article
- Monitoring of circulating tumour DNA in advanced pancreatic ductal adenocarcinoma predicts clinical outcome and reveals disease progression earlier than radiological imaging.Molecular oncology · 2023Article
- An enzymatic on/off switch-mediated assay for KRAS hotspot point mutation detection of circulating tumor DNA.Journal of clinical laboratory analysis · 2020Article
- PNA Clamping in Nucleic Acid Amplification Protocols to Detect Single Nucleotide Mutations Related to Cancer.Molecules (Basel, Switzerland) · 2020Review
- Mutation analysis by deep sequencing of pancreatic juice from patients with pancreatic ductal adenocarcinoma.BMC cancer · 2019Article
- 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
- Kinetic hairpin oligonucleotide blockers for selective amplification of rare mutations.Scientific reports · 2014Article
- Influence of microsatellite instability and KRAS and BRAF mutations on lymph node harvest in stage I-III colon cancers.Molecular medicine (Cambridge, Mass.) · 2013Article
- Discovery and characterization of artifactual mutations in deep coverage targeted capture sequencing data due to oxidative DNA damage during sample preparation.Nucleic acids research · 2013Article
- Detecting KRAS mutations in peripheral blood of colorectal cancer patients by peptide nucleic acid clamp PCR.Experimental and therapeutic medicine · 2012Article
- Heterogeneous distribution of K-ras mutations in primary colon carcinomas: implications for EGFR-directed therapy.International journal of colorectal disease · 2011Article
- Phylogenetic characterisation of naturally occurring feline immunodeficiency virus in the United Kingdom.Veterinary microbiology · 2011Article
- Molecular diagnosis of response to neoadjuvant chemoradiation therapy in patients with locally advanced rectal cancer.Journal of the American College of Surgeons · 2011Article
- SNaPshot and StripAssay as valuable alternatives to direct sequencing for KRAS mutation detection in colon cancer routine diagnostics.The Journal of molecular diagnostics : JMD · 2011Article
- Detection of exon 12 Mutations in the JAK2 gene: enhanced analytical sensitivity using clamped PCR and nucleotide sequencing.The Journal of molecular diagnostics : JMD · 2010Article
Corrections and comments
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Sensitive detection of tumor-specific point mutations is of interest in both the early detection of cancer and the monitoring of treatment at a molecular level. Recently, peptide nucleic acid (PNA) clamp real-time PCR has provided a time-sparing and sensitive method for the detection of mutations in the presence of a large excess of wild-type DNA. We present the first report that the sensitivity of PNA clamp PCR is limited by the low fidelity of TaqDNA polymerase. Replication errors introduced by Taq polymerase in the PNA-binding site were amplified during PCR due to the resulting mismatches between PNA and DNA. To reduce the frequency of polymerase-induced errors, we developed a PNA clamp PCR assay for the detection of mutations in codons 12 and 13 of the K-ras gene based on a high-fidelity DNA polymerase. The sensitivity of our assay increased approximately 10-fold, significantly detecting mutant DNA diluted 20,000-fold in wild-type DNA (P = 0.025), compared with its detection at 2000-fold dilution (P = 0.039) when Taq polymerase was used. Our data suggest that the replication errors caused by Taq polymerase must be taken into consideration for PNA clamp PCR and for other methods based on selective PCR amplification, and that these assays can be enhanced by high-fidelity DNA polymerases.
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