title: Comparison of molecular testing methods for the detection of EGFR mutations in formalin-fixed paraffin-embedded tissue specimens of non-small cell lung cancer.
abstract: OBJECTIVE
  To conduct a methods correlation study of three different assays for the detection of mutations at EGFR gene in human formalin-fixed paraffin-embedded tumour (FFPET) specimens of non-small cell lung carcinomas (NSCLC).
  
  
  METHODS
  We conducted a 2-site method comparison study of two european conformity (CE) in vitro diagnostic (IVD)-marked assays, the cobas EGFR Mutation Test and the Therascreen EGFR29 Mutation Kit, and 2× bidirectional Sanger sequencing. We blind-tested 124 NSCLC FFPET specimens with all three methods; the cobas test was performed at both sites. Positive (PPA) and negative percent agreements (NPA) were determined for the cobas test versus each of the other two methods. Specimens yielding discordant test results between methods were further tested using quantitative massively parallel pyrosequencing (MPP).
  
  
  RESULTS
  PPA between cobas and Sanger was 98.8%; NPA was 79.3%. Overall there were seven discordant results. MPP confirmed an exon 19 deletion in two cases and L858R mutation in four cases. PPA between cobas and Therascreen was 98.9% and NPA was 100%. There was one discordant result. Reproducibility of the cobas test between the two sites was 99.2%.
  
  
  CONCLUSIONS
  The invalid rates for the cobas test and Therascreen were lower than Sanger sequencing. The cobas and Therascreen assays showed a high degree of concordance, and both were more sensitive for the detection of exon 19 deletion and L858R mutations than Sanger. The cobas test was highly reproducible between the two testing sites, used the least amount of DNA input and was the only test with automated results reporting.
authors: Laboratorio de Dianas Terapeuticas, Centro Integral Oncologico Clara Campal, Hospital Universitario Madrid Sanchinarro, Madrid, Spain. flopezrios@hospitaldemadrid.com
authors: Fernando
authors: F
authors: Lopez-Rios
authors: Barbara
authors: B
authors: Angulo
authors: Belen
authors: B
authors: Gomez
authors: Debbie
authors: D
authors: Mair
authors: Rebeca
authors: R
authors: Martinez
authors: Esther
authors: E
authors: Conde
authors: Felice
authors: F
authors: Shieh
authors: Julie
authors: J
authors: Tsai
authors: Jeffrey
authors: J
authors: Vaks
authors: Robert
authors: R
authors: Current
authors: H Jeffrey
authors: HJ
authors: Lawrence
authors: David
authors: D
authors: Gonzalez de Castro
mesh_terms: D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics*; D005557:Formaldehyde; D018773:Genes, erbB-1 / Q000235:genetics*; D005821:Genetic Techniques; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics*; D009154:Mutation; D016612:Paraffin Embedding; D015203:Reproducibility of Results; D016707:Tissue Fixation
chemical_list: D005557:Formaldehyde
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