ArticlePloS one2012
Allele specific locked nucleic acid quantitative PCR (ASLNAqPCR): an accurate and cost-effective assay to diagnose and quantify KRAS and BRAF mutation.
Article in PloS one, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 28 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
28 citing papers in PubMed.
- Expression of 19 microRNAs in glioblastoma and comparison with other brain neoplasia of grades I-III.Molecular oncology · 2014Trial
- KRAS mutations detection methodology: from RFLP to CRISPR/Cas based methods.Functional & integrative genomics · 2024Review
- High throughput AS LNA qPCR method for the detection of a specific mutation in poliovirus vaccine strains.Vaccine · 2024Article
- Combining the amplification refractory mutation system and high-resolution melting analysis for KRAS mutation detection in clinical samples.Analytical and bioanalytical chemistry · 2023Article
- Metastatic Melanoma: Liquid Biopsy as a New Precision Medicine Approach.International journal of molecular sciences · 2023Review
- Molecular testing on serous effusion: An update.CytoJournal · 2021Review
- From Neurosurgical Planning to Histopathological Brain Tumor Characterization: Potentialities of Arcuate Fasciculus Along-Tract Diffusion Tensor Imaging Tractography Measures.Frontiers in neurology · 2021Article
- Isolation of extracellular vesicles improves the detection of mutant DNA from plasma of metastatic melanoma patients.Scientific reports · 2020Article
- Targeting Endogenous K-RAS for Degradation through the Affinity-Directed Protein Missile System.Cell chemical biology · 2020Article
- Evaluation of the expression levels of BRAFBMC cancer · 2020Article
- Review
- Article
- Allele-Specific Isothermal Amplification Method Using Unmodified Self-Stabilizing Competitive Primers.Analytical chemistry · 2018Article
- Establishing PNB-qPCR for quantifying minimal ctDNA concentrations during tumour resection.Scientific reports · 2017Article
- Methylation of ZNF331 is an independent prognostic marker of colorectal cancer and promotes colorectal cancer growth.Clinical epigenetics · 2017Article
- Sensitive High-Resolution Melting Analysis for Screening of KRAS and BRAF Mutations in Iranian Human Metastatic Colorectal CancersAsian Pacific journal of cancer prevention : APJCP · 2016Article
- Braf, Kras and Helicobacter pylori epigenetic changes-associated chronic gastritis in Egyptian patients with and without gastric cancer.World journal of microbiology & biotechnology · 2016Article
- Detection of Rare Drug Resistance Mutations by Digital PCR in a Human Influenza A Virus Model System and Clinical Samples.Journal of clinical microbiology · 2016Article
- Article
- BRAF V600E and risk stratification of thyroid microcarcinoma: a multicenter pathological and clinical study.Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc · 2015Article
Corrections and comments
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Authors and funding
8 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The use of tyrosine kinase inhibitors (TKIs) requires the testing for hot spot mutations of the molecular effectors downstream the membrane-bound tyrosine kinases since their wild type status is expected for response to TKI therapy. We report a novel assay that we have called Allele Specific Locked Nucleic Acid quantitative PCR (ASLNAqPCR). The assay uses LNA-modified allele specific primers and LNA-modified beacon probes to increase sensitivity, specificity and to accurately quantify mutations. We designed primers specific for codon 12/13 KRAS mutations and BRAF V600E, and validated the assay with 300 routine samples from a variety of sources, including cytology specimens. All were analyzed by ASLNAqPCR and Sanger sequencing. Discordant cases were pyrosequenced. ASLNAqPCR correctly identified BRAF and KRAS mutations in all discordant cases and all had a mutated/wild type DNA ratio below the analytical sensitivity of the Sanger method. ASLNAqPCR was 100% specific with greater accuracy, positive and negative predictive values compared with Sanger sequencing. The analytical sensitivity of ASLNAqPCR is 0.1%, allowing quantification of mutated DNA in small neoplastic cell clones. ASLNAqPCR can be performed in any laboratory with real-time PCR equipment, is very cost-effective and can easily be adapted to detect hot spot mutations in other oncogenes.
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