Wobble-enhanced ARMS Method for Detection of KRAS and BRAF Mutations

Hamfjord, Julian; Stangeland, Astrid M.; Skrede, Martina L.; Ikdahl, Tone; Kure, Elin H.

doi:10.1097/pdm.0b013e31820b49e2

Cited by 22 publications

(16 citation statements)

References 14 publications

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“…Input in the KRAS and NRAS assays were 10 ng and 20 ng DNA, respectively. Analyses for the BRAF V600E mutation was performed as described by Hamfjord et al [48]…”

Section: Methodsmentioning

confidence: 99%

Interleukin-6 and C-reactive protein as prognostic biomarkers in metastatic colorectal cancer

et al. 2016

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ObjectivesThe aim was to explore the prognostic significance of IL-6 and markers of systemic inflammatory response (SIR), in particular C-reactive protein (CRP), in metastatic colorectal cancer (mCRC) patients, in the total study population and according to RAS and BRAF mutation status.ResultsHigh levels of pretreatment serum IL-6 or CRP were associated with impaired outcome, in terms of reduced PFS and OS. Patients with low versus high serum IL-6 levels had median OS of 26.0 versus 16.6 months, respectively (P < 0.001). Stratified according to increasing CRP levels, median OS varied from 24.3 months to 12.3 months, (P < 0.001). IL-6 and CRP levels affected overall prognosis also in adjusted analyses. The effect of IL-6 was particularly pronounced in patients with BRAF mutation (interaction P = 0.004).Materials and MethodsIL-6 and CRP were determined in pre-treatment serum samples from 393 patients included in the NORDIC-VII trial, in which patients with mCRC received first line treatment. The effect of serum IL-6 and CRP on progression-free survival (PFS) and overall survival (OS) was estimated.ConclusionsHigh baseline serum consentrations of IL-6 or CRP were associated with impaired prognosis in mCRC. IL-6 and CRP give independent prognostic information in addition to RAS and BRAF mutation status.

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“…Input in the KRAS and NRAS assays were 10 ng and 20 ng DNA, respectively. Analyses for the BRAF V600E mutation was performed as described by Hamfjord et al [48]…”

Section: Methodsmentioning

confidence: 99%

Interleukin-6 and C-reactive protein as prognostic biomarkers in metastatic colorectal cancer

et al. 2016

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“…THxID™-BRAF used an ARMS based PCR technology. ARMS PCR approaches have been widely and successfully used to detect somatic mutation in routine research, notably BRAF mutations [16,29,30]. One of the advantages of ARMS-PCR is that the assay is designed to amplify a relative larger common fragment of DNA that flanks the mutation site in all samples regardless of their mutation status.…”

Section: Discussionmentioning

confidence: 99%

Comparative evaluation of the new FDA approved THxID™-BRAF test with high resolution melting and sanger sequencing

et al. 2014

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BackgroundSince patients diagnosed with BRAF V600E and V600K mutated advanced melanoma show response to treatment with MAP kinase inhibitors, several sensitive methods have been developed to determine the V600 allele status of melanoma patients. Vemurafenib (Zelboraf) and dabrafenib (Tafinlar) are specific BRAF V600 inhibitors recently approved by the US FDA as single agent treatments for unresectable or metastatic melanoma in patients with the BRAF V600 mutation.MethodsWe assessed the new CE THxID™-BRAF diagnostic test, which is also FDA-approved as a companion diagnostic test in the US under a specific reference and compared the results of this assay with both High Resolution Melting (HRM) and Sanger sequencing in 113 melanoma FFPE samples.ResultsInvalid results were observed in 0/113 specimen with HRM, 5/113 (4.4%) with Sanger sequencing, and 1/113 (0.9%) with the THxID™-BRAF test. Positive percentage agreement (PPA) was 93.5% (95% CI 82.5 - 97.8) for V600E and V600K mutations combined for the THxID™-BRAF test and HRM, and negative percentage agreement (NPA) was 100.0% (95% CI 94.5 - 100.0). For the THxID™-BRAF test and Sanger, PPA was 100.0% (95% CI 92.1 - 100.0) and NPA 100.0% (95% CI 94.2 - 100.0). One V600E sample identified by THxID™-BRAF test was detected as wild-type by HRM and uninterpretable by Sanger. All V600K (n = 3) were detected using the 3 different approaches. Finally, percent agreement values were not significantly different when using punches (n = 77) vs. slides (n = 36) or depending on samples characteristics such as pigmentation, necrosis, and tumor content.ConclusionsThis study demonstrated the high agreement between the FDA approved THxID™-BRAF assay, HRM, and Sanger sequencing. It has also highlighted the potential of THxID™-BRAF to be applied to a broader range of sample types than claimed in the current “instructions for use”, an extension that would require the ad hoc validation and approval.

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“…The KRAS mutations (codons 12 and 13) were determined using the Wobble-enhanced amplification refractory mutation system [11].…”

Section: Methodsmentioning

confidence: 99%

Generation and Characterisation of Novel Pancreatic Adenocarcinoma Xenograft Models and Corresponding Primary Cell Lines

et al. 2014

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Pancreatic adenocarcinoma is one of the most lethal cancer types, currently lacking efficient treatment. The heterogeneous nature of these tumours are poorly represented by the classical pancreatic cell lines, which have been through strong clonal selection in vitro, and are often derived from metastases. Here, we describe the establishment of novel pancreatic adenocarcinoma models, xenografts and corresponding in vitro cell lines, from primary pancreatic tumours. The morphology, differentiation grade and gene expression pattern of the xenografts resemble the original tumours well. The cell lines were analysed for colony forming capacity, tumourigenicity and expression of known cancer cell surface markers and cancer stem-like characteristics. These primary cell models will be valuable tools for biological and preclinical studies for this devastating disease.

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Wobble-enhanced ARMS Method for Detection of KRAS and BRAF Mutations

Cited by 22 publications

References 14 publications

Interleukin-6 and C-reactive protein as prognostic biomarkers in metastatic colorectal cancer

Interleukin-6 and C-reactive protein as prognostic biomarkers in metastatic colorectal cancer

Comparative evaluation of the new FDA approved THxID™-BRAF test with high resolution melting and sanger sequencing

Generation and Characterisation of Novel Pancreatic Adenocarcinoma Xenograft Models and Corresponding Primary Cell Lines

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