Competitive allele specific TaqMan PCR for KRAS, BRAF and EGFR mutation detection in clinical formalin fixed paraffin embedded samples

2012 ◽  
Vol 92 (3) ◽  
pp. 275-280 ◽  
Author(s):  
Audrey Didelot ◽  
Delphine Le Corre ◽  
Armelle Luscan ◽  
Aurélie Cazes ◽  
Karine Pallier ◽  
...  
Keyword(s):  
Mutation Detection ◽  
Egfr Mutation ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Allele Specific ◽  
Taqman Pcr ◽  
Formalin Fixed

scholarly journals Droplet Digital PCR for Mutation Detection in Formalin-Fixed, Paraffin-Embedded Melanoma Tissues

2018 ◽  
Vol 20 (2) ◽  
pp. 240-252 ◽  
Author(s):  
Ashleigh C. McEvoy ◽  
Benjamin A. Wood ◽  
Nima M. Ardakani ◽  
Michelle R. Pereira ◽  
Robert Pearce ◽  
...  
Keyword(s):  
Mutation Detection ◽  
Digital Pcr ◽  
Droplet Digital Pcr ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed

scholarly journals Analysis ofKRASMutations of Exon 2 Codons 12 and 13 by SNaPshot Analysis in Comparison to Common DNA Sequencing

2010 ◽  
Vol 2010 ◽  
pp. 1-5 ◽  
Author(s):  
Rica Zinsky ◽  
Servet Bölükbas ◽  
Holger Bartsch ◽  
Joachim Schirren ◽  
Annette Fisseler-Eckhoff
Keyword(s):  
Dna Sequencing ◽  
Mutation Detection ◽  
Nonsmall Cell Lung Cancer ◽  
Tissue Samples ◽  
Exon 2 ◽  
Mutation Status ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Status Analysis ◽  
Formalin Fixed

Due to the call for fastKRASmutation status analysis for treatment of patients with monoclonal antibodies for metastatic colorectal cancer, sensitive, economic, and easily feasible methods are required. Under this aspect, the sensitivity and specificity of the SNaPshot analysis in comparison to the commonly used DNA sequencing was checked. We examinedKRASmutations in exon 2 codons 12 and 13 with DNA sequencing and SNaPshot analysis in 100 formalin-fixed paraffin-embedded tumor tissue samples of pancreatic carcinoma, colorectal carcinoma, and nonsmall cell lung cancer specimens of the primary tumor or metastases. 40% of these samples demonstrated mutatedKRASgenes using sequencing and SNaPshot-analysis; additional five samples (45/100) were identified only with the SNaPshot.KRASmutation detection is feasible with the reliable SNaPshot analysis method. The more frequent mutation detection by the SNaPshot analysis shows that this method has a high probability of accuracy in the detection ofKRASmutations compared to sequencing.

scholarly journals KRAS Mutation Detection in Paired Frozen and Formalin-Fixed Paraffin-Embedded (FFPE) Colorectal Cancer Tissues

2011 ◽  
Vol 12 (5) ◽  
pp. 3191-3204 ◽  
Author(s):  
Jérome Solassol ◽  
Jeanne Ramos ◽  
Evelyne Crapez ◽  
Majda Saifi ◽  
Alain Mangé ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Kras Mutation ◽  
Mutation Detection ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Cancer Tissues ◽  
Formalin Fixed

scholarly journals Understanding the Pattern of Oropharyngeal Cancers from North-East Romanian Patients

2021 ◽  
Vol 11 (24) ◽  
pp. 12079
Author(s):  
Ramona Gabriela Ursu ◽  
Simona Eliza Giusca ◽  
Irene Alexandra Spiridon ◽  
Bianca Manole ◽  
Mihai Danciu ◽  
...  
Keyword(s):  
Tongue Cancer ◽  
Growth Factor Receptor ◽  
Hpv Dna ◽  
North East ◽  
Formalin Fixed Paraffin ◽  
Screening Strategies ◽  
Formalin Fixed Paraffin Embedded ◽  
Allele Specific ◽  
Hpv18 E6 ◽  
Taqman Pcr

Background: Human papilloma virus (HPV) is acknowledged as a risk factor for oropharyngeal squamous cellular cancers (OPSCC), of which the dominant types are tonsillar (TSCC) and base of tongue cancer (BOTSCC). Objective: To assess the role of HPV in selected OPSCC cases, from Romanian patients by sensitive and complementary molecular assays. Material and Methods: Fifty-four formalin fixed paraffin embedded (FFPE) OPSCC samples were analyzed for HPV DNA by a PCR-based bead-based multiplex-assay. Thirty-four samples were tested for HPV RNA and for overexpression of p16INK4a by immunohistochemistry. Twenty samples were evaluated by Competitive Allele-Specific Taqman PCR (CAST-PCR) for fibroblast growth factor receptor 3 protein (FGFR3) status. Results: A total of 33.3% (18/54) OPSCC samples were positive for HPV DNA. HPV16 was the most frequent type (30%, 16/54); followed by HPV18 (3.7%, 2/54); and 1 sample (1.8%) was positive for both HPV16 and 18. HPV18 E6*I was detected in a HPV18 DNA-positive oropharynx tumor. Four samples positive for HPV16 were also positive for p16INK4a. All the tested samples were negative for FGFR3. Conclusions: The increased HPV16 prevalence is in line with similar studies and is a new confirmation that HPV16 is the most prevalent type in our country; supporting the potential benefit of prophylactic vaccines. Overall, there is no concordance between DNA and any of the two other analytes that are considered being markers of HPV-driven cancers. There is a need to explore novel screening strategies that could be broadly used in the clinical routine to initiate preventive measures.

scholarly journals Clinical Performance Evaluation Of The Idylla™ Egfr Mutation Test On Formalin-Fixed Paraffin-Embedded Tissue Of Non-Small Cell Lung Cancer

2020 ◽  
Author(s):  
Mercedes Delgado-Garcia ◽  
Birgit Weynand ◽  
Lourdes Gómez Izquierdo ◽  
María José Hernández Barrera ◽  
Ángela María Blanco Lobo ◽  
...  
Keyword(s):  
Egfr Mutation ◽  
Small Cell ◽  
Test Method ◽  
Egfr Mutations ◽  
Small Cell Lung ◽  
Diagnostic Agreement ◽  
Mutational Status ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed

Abstract Background detection of epidermal growth factor receptor ( EGFR) mutations in exons 18-21 is recommended in all patients with advanced Non-small-cell lung carcinoma due to the demonstrated efficiency of the standard therapy with tyrosine kinase inhibitors in EGFR mutated patients. Therefore, choosing a suitable technique to test EGFR mutational status is crucial to warrant a valid result in a short turnaround time using the lowest possible amount of tissue material. The Idylla™ EGFR Mutation Test is a simple, fast and reliable method designed for the detection of EGFR mutations from formalin-fixed paraffin-embedded samples. The aim of this study was the Clinical Performace Evaluation of the Idylla™ EGFR Mutation Test on the Idylla™ System. Methods EGFR mutational status was determined on 132 archived formalin-fixed paraffin-embedded tissue sections with Idylla™ technology. Results were compared with the results previously obtained by routine method in the reference lab (Therascreen ® EGFR RGQ PCR v2, Qiagen in Molecular Pathology lab, Hospital Universitario Virgen del Rocío de Sevilla). Results the overall agreement between results obtained with the Idylla™ EGFR Mutation Test and the Comparator test method was 95.38% (with 1-sided 95% lower limit of 91.7%) showing Positive Diagnostic Agreement of 93.22% and Negative Diagnostic Agreement of 97.18%, with a Limit Of Detection ≤5%. Conclusions the Idylla™ EGFR Mutation Test passed its clinical validity performance characteristics for accuracy.

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