Detection of EGFR and BRAF mutations by competitive allele-specific TaqMan polymerase chain reaction in lung adenocarcinoma

Yang, Yang; Yue, Meng; Zhang, Hang; Shen, Xizhong; Li, Rutian; Yu, Lan; Liu, Baorui; Wang, Lifeng

doi:10.3892/ol.2017.7652

Cited by 8 publications

(6 citation statements)

References 87 publications

(85 reference statements)

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“…The BRAF gene is mutated in 1–5% of NSCLC cases and most of these mutations occur in adenocarcinoma (14). BRAF is a member of the RAF family of serine/threonine kinases that mediate signal transduction between RAS and mitogen-activated protein kinase (MAPK) signaling pathways (15).…”

Section: Discussionmentioning

confidence: 99%

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Multiple primary malignant neoplasms: A case report and literature review

et al. 2019

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Until recently, few cases of three or more malignant tumors in one patient have been reported. Owing to the high incidence rate of these tumors, the improvement in cancer diagnosis and treatment, and the extension of patient survival time, the incidence of reported multiple primary malignant neoplasms has gradually increased. The present study reported the case of a 57-year-old man with non-small cell lung cancer combined with B-Raf proto-oncogene serine/threonine kinase V600E mutation, gastrointestinal stromal tumors and lumbar vertebral malignant mucinous sarcoma. The pathogenesis, diagnosis and treatment of these three malignancies are discussed and previous studies are also reviewed. The aim of the study was to analyze the genetic mutations associated with multiple primary malignant tumors and to discuss whether those mutations with unknown functional significance could be used as therapeutic indicators. This case report will serve as a reference for future treatment of such patients.

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Section: Discussionmentioning

confidence: 99%

“…9) (16). The V600E mutation accounts for ~50% of BRAF mutations (14). In a multicenter, single arm, non-randomized phase II study (BRF113928; identifier, NCT01336634) (17), the efficacy of dabrafenib as a monotherapy was compared with that of dabrafenib administered in combination with trametinib.…”

Section: Discussionmentioning

confidence: 99%

Multiple primary malignant neoplasms: A case report and literature review

et al. 2019

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“…In a first set of experiments, we compared three widely used methods to determine levels of mutated BRAF: an allele-specific PCR method, 7 a commercially available Taqman Mutation Detection Assay which was already used in several studies, 10,16 and a commercially available digital droplet PCR Kit. 17 To evaluate the three methods, we diluted genomic DNA from the A673 cell line, which contains a heterozygous BRAFV600E mutation with wildtype DNA extracted from blood of healthy donors ranging from 100% mutated cells to 0.01% mutated cells and performed the PCR reactions in parallel.…”

Section: Resultsmentioning

confidence: 99%

Longitudinal assessment of peripheral blood BRAFV600E levels in patients with Langerhans cell histiocytosis

et al. 2018

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BACKGROUND: Langerhans cell histiocytosis (LCH) is a histiocytic disorder driven by a constitutive activation of the MAPK signaling pathway in myeloid cells. In 50-60% of cases, it is caused by the BRAFV600E mutation. There is evidence that levels of BRAFV600E in the peripheral blood of patients with LCH correlate with disease burden and could be used as marker for disease extent and response to therapy. However, there is currently no consensus on how testing for minimal disseminated disease should be performed. METHODS: Different approaches to determine the mutation load in patients with LCH were assessed and longitudinal evaluation of patient DNA during treatment with chemotherapy and/or the RAF inhibitor vemurafenib was performed. DNA was isolated from whole blood, different leukocyte subsets, and circulating cell-free DNA (ccf-DNA). RESULTS: We show that determining BRAF levels from whole blood is superior to using ccfDNA. Furthermore, it is important to identify the clinically relevant BRAF-mutated cellular subpopulations such as CD14 + monocytes or CD1c + DCs, since other blood cells can also harbor the mutation and therefore confound whole blood or ccfDNA measurements. CONCLUSION: Our data support the view that single-agent treatment with an RAF inhibitor reduces disease activity but does not cure LCH.

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“…In addition, enhanced allele-specific qPCR approaches have been described with improved sensitivity compared to ARMS. For example, competitive allele-specific TaqMan PCR has a reported LoD of 0.1% for EGFR L858R and 1% for T790M 24 . Similarly, modified blocking oligonucleotides have been applied to reduce the LoD to 0.1% for four common KRAS variants 25 .…”

Section: Discussionmentioning

confidence: 99%

Single-copy detection of somatic variants from solid and liquid biopsy

Silva¹,

Powalowska²,

Stolarek³

et al. 2021

Sci Rep

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Accurate detection of somatic variants, against a background of wild-type molecules, is essential for clinical decision making in oncology. Existing approaches, such as allele-specific real-time PCR, are typically limited to a single target gene and lack sensitivity. Alternatively, next-generation sequencing methods suffer from slow turnaround time, high costs, and are complex to implement, typically limiting them to single-site use. Here, we report a method, which we term Allele-Specific PYrophosphorolysis Reaction (ASPYRE), for high sensitivity detection of panels of somatic variants. ASPYRE has a simple workflow and is compatible with standard molecular biology reagents and real-time PCR instruments. We show that ASPYRE has single molecule sensitivity and is tolerant of DNA extracted from plasma and formalin fixed paraffin embedded (FFPE) samples. We also demonstrate two multiplex panels, including one for detection of 47 EGFR variants. ASPYRE presents an effective and accessible method that simplifies highly sensitive and multiplexed detection of somatic variants.

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Detection of EGFR and BRAF mutations by competitive allele-specific TaqMan polymerase chain reaction in lung adenocarcinoma

Cited by 8 publications

References 87 publications

Multiple primary malignant neoplasms: A case report and literature review

Multiple primary malignant neoplasms: A case report and literature review

Longitudinal assessment of peripheral blood BRAFV600E levels in patients with Langerhans cell histiocytosis

Single-copy detection of somatic variants from solid and liquid biopsy

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