Pyrosequencing for EGFR Mutation Detection

2013 ◽  
Vol 22 (4) ◽  
pp. 196-203 ◽  
Author(s):  
Nora Sahnane ◽  
Rossana Gueli ◽  
Maria G. Tibiletti ◽  
Barbara Bernasconi ◽  
Michele Stefanoli ◽  
...  
Keyword(s):  
Mutation Detection ◽  
Egfr Mutation

scholarly journals EGFR mutation detection in circulating cell-free DNA of lung adenocarcinoma patients: analysis of LUX-Lung 3 and 6

2016 ◽  
Vol 116 (2) ◽  
pp. 175-185 ◽  
Author(s):  
Yi-Long Wu ◽  
Lecia V Sequist ◽  
Cheng-Ping Hu ◽  
Jifeng Feng ◽  
Shun Lu ◽  
...  
Keyword(s):  
Lung Adenocarcinoma ◽  
Mutation Detection ◽  
Egfr Mutation ◽  
Cell Free Dna ◽  
Free Dna ◽  
Circulating Cell Free Dna

scholarly journals T790M EGFR Mutation Detection in Cerebrospinal Fluid and Response to Osimertinib in a Lung Cancer Patient with Meningeal Carcinomatosis

2017 ◽  
Vol 12 (9) ◽  
pp. e138-e139 ◽  
Author(s):  
Hugo Gortais ◽  
Catherine Daniel ◽  
François-Clément Bidard ◽  
Emmanuelle Jeannot ◽  
Céline Callens ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Cerebrospinal Fluid ◽  
Cancer Patient ◽  
Lung Cancer Patient ◽  
Mutation Detection ◽  
Egfr Mutation ◽  
Meningeal Carcinomatosis

scholarly journals Clinical Validation of KRAS, BRAF, and EGFR Mutation Detection Using Next-Generation Sequencing

2014 ◽  
Vol 141 (6) ◽  
pp. 856-866 ◽  
Author(s):  
Ming-Tseh Lin ◽  
Stacy L. Mosier ◽  
Michele Thiess ◽  
Katie F. Beierl ◽  
Marija Debeljak ◽  
...  
Keyword(s):  
Next Generation Sequencing ◽  
Mutation Detection ◽  
Egfr Mutation ◽  
Clinical Validation ◽  
Next Generation ◽  
Generation Sequencing

scholarly journals Plasma EGFR Mutation Detection Associated With Survival Outcomes in Advanced-Stage Lung Cancer

2015 ◽  
Vol 16 (6) ◽  
pp. 507-513 ◽  
Author(s):  
David C.L. Lam ◽  
Terence C.C. Tam ◽  
Kenneth M.K. Lau ◽  
Wai-Mui Wong ◽  
Christopher K.M. Hui ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Mutation Detection ◽  
Egfr Mutation ◽  
Survival Outcomes ◽  
Advanced Stage ◽  
Advanced Stage Lung Cancer ◽  
Stage Lung Cancer

Detection of clinically significant mutations in the epidermal growth factor receptor missed by direct sequencing using a highly sensitive DNA endonuclease

2006 ◽  
Vol 24 (18_suppl) ◽  
pp. 10054-10054
Author(s):  
A. M. Borras ◽  
Y. Kuang ◽  
A. M. Rogers ◽  
A. J. Holmes ◽  
M. Gallegos Ruiz ◽  
...  
Keyword(s):  
Epidermal Growth Factor Receptor ◽  
Mutation Detection ◽  
Egfr Mutation ◽  
Direct Sequencing ◽  
Growth Factor Receptor ◽  
Egfr Mutations ◽  
Dna Endonuclease ◽  
Epidermal Growth ◽  
Clinically Significant ◽  
Sensitive Method

10054 Background: Mutations in the epidermal growth factor receptor (EGFR) are associated with sensitivity and resistance to EGFR inhibitors gefitinib and erlotinib in patients with non-small cell lung carcinoma (NSCLC). Direct sequencing is currently used for mutation detection but sensitivity is limited and requires dissection to obtain a relatively pure population of tumor cells. We examined a DNA endonuclease, SURVEYOR, which cleaves mismatched heteroduplexed DNA, as a more sensitive method for EGFR mutation screening and compared it to direct sequencing. Methods: EGFR exons 18–21 from tumor DNA were amplified using PCR, digested with SURVEYOR, and the products analyzed by HPLC. Specimens that produced digestion products were re-analyzed by size separation or by denaturing HPLC followed by fractionation and sequencing. Tumor specimens from 191 NSCLC patients were analyzed: 61 frozen tumors specimens; 91 dissected formalin fixed paraffin embedded (FFPE) and 39 un-dissected FFPE tumor specimens from patients treated with gefitinib or erlotinib in whom clinical outcome was available. 173 specimens were independently analyzed by direct sequencing. Results: We detected 48 EGFR mutations by sequencing and 61 using SURVEYOR. All EGFR mutations identified by sequencing, including those using un-dissected tumor specimens, were detected by SURVEYOR and none were missed (sensitivity: 100%, negative predictive value: 100%). 13 mutations were detected by SURVEYOR not detected by sequencing. This included 5 mutations (4 exon 19 deletions; 1 L858R) in 7 (71%) patients who clinically had a PR to gefitinib or erlotinib but who were wild type by sequencing. In 4 patients, 2 with clinical acquired resistance to gefitinib, a T790M mutation was found which was undetected by sequencing. Conclusions: SURVEYOR analysis is a more sensitive method for EGFR mutation detection than direct sequencing. It can be used to detect EGFR mutations from un-dissected tumor specimens and can detect clinically significant activating or resistance associated EGFR mutations not detected by direct sequencing. [Table: see text]

EGFR mutation detection on routine cytological smears of non-small cell lung cancer by digital PCR: a validation study

2016 ◽  
Vol 69 (5) ◽  
pp. 454-457 ◽  
Author(s):  
Umberto Malapelle ◽  
Caterina de Luca ◽  
Elena Vigliar ◽  
Francesca Ambrosio ◽  
Danilo Rocco ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Mutation Detection ◽  
Egfr Mutation ◽  
Growth Factor Receptor ◽  
Digital Pcr ◽  
Neoplastic Cell ◽  
Egfr Mutations ◽  
Taqman Assay ◽  
Cell Percentage ◽  
Egfr Mutant

Highly sensitive genotyping techniques are useful to detect epidermal growth factor receptor (EGFR) mutations on lung cancer cytological samples, when these specimens feature only few neoplastic cells. This study aimed to validate digital PCR (dPCR) methodology on cytological material. In plasmid model system, dPCR allowed for the detection of a minimal percentage (1%) of EGFR mutant alleles. Cytological samples (n=30), with neoplastic cell percentage ranging from 10% to 80% and yielding a quantity of extracted DNA ranging from 1.75 to 60 ng/µL were selected. Results previously generated by fragment length and TaqMan assays (n=8 exon 19 deletions, n=2 L858R mutations and n=20 wild-type DNA) were compared with those obtained by dPCR. Data were highly concordant (96.6%). However, dPCR detected an additional L858R mutation that had been missed by TaqMan assay on a paucicellular smear. This mutation was confirmed by cloning PCR products and sequencing. Thus, dPCR can reliably be used to increase EGFR mutation detection rate on scarcely cellular lung cancer smears.

scholarly journals The diagnostic accuracy of pleural effusion and plasma samples versus tumour tissue for detection of EGFR mutation in patients with advanced non-small cell lung cancer: comparison of methodologies

2013 ◽  
Vol 66 (12) ◽  
pp. 1065-1069 ◽  
Author(s):  
Xiaoqing Liu ◽  
Yachao Lu ◽  
Guanshan Zhu ◽  
Yao Lei ◽  
Li Zheng ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Pleural Effusion ◽  
Small Cell Lung Cancer ◽  
Sanger Sequencing ◽  
Mutation Detection ◽  
Egfr Mutation ◽  
Small Cell ◽  
Plasma Samples ◽  
Cell Block ◽  
Small Cell Lung

AimsTo evaluate the suitability of malignant pleural effusion (MPE) and plasma as surrogate samples for epidermal growth factor receptor (EGFR) mutation detection, and compare three different detection methods.MethodsMatched tissue and plasma samples were collected from patients with advanced non-small cell lung cancer (NSCLC) (stage IIIB/IV adenocarcinoma/adenosquamous carcinoma), with matched MPE samples collected from a subgroup. DNA was extracted from tissue, MPE cell block, MPE supernatant and plasma before mutation detection by amplification refractory mutation system (ARMS) (all samples), Sanger sequencing and mutant-specific immunohistochemistry (IHC) (tissue and MPE cell blocks only).ResultsSensitivity of MPE cell block, MPE supernatant and plasma versus tissue: 81.8% (9/11), 63.6% (7/11) and 67.5% (27/40); specificity was 80.0% (8/10), 100% (10/10) and 100% (46/46), respectively. Sensitivity of Sanger sequencing versus ARMS: 81.8% (27/33) for tissue, 40% (4/10) for MPE cell blocks; specificity was 100% (36/36 and 12/12) for both. Sensitivity of mutant-specific IHC versus ARMS: 54.8% (17/31) for tissue, 50.0% (6/12) for MPE cell blocks; specificity was 97.1% (34/35) and 100% (14/14), respectively.ConclusionsMPE and plasma are valid surrogates for NSCLC tumour EGFR mutation detection when tissue is not available. ARMS is most suitable for mutation detection in tissue and MPE cell blocks; however, mutant-specific IHC could be a complementary method when DNA-based molecular testing is unavailable.

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
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