title: [Validation steps for the detection of L858R mutation in EGFR gene by real time quantitative PCR].
abstract: Since January 16(th) 2010, the French legislation requires that the medical laboratories must be accredited according to ISO 15189 standards. This concerned all the biological medical technics, including molecular biology technics. In this work, we described the validation steps by real time quantitative PCR of L858R mutation in EGFR gene, frequently detected in non-small lung cancers (NSCLC). Epidermal growth factor - tyrosine kinase inhibitors (EGFR-TKIs) are authorized in Europe for the treatment of metastatic NSCLC after failure of, at least one, prior chemotherapy. Thus, in view of accreditation of this analysis, we have used the recommendation of the COFRAC (Comité français d'accréditation) and INCa (Institut national du cancer). Several parameters have been tested, such as the primers, the limit of detection, and the sensitivity and specificity of the method. In addition, a risk study has been evaluated. Although long and fastidious, the method of validation is required to perform analysis in optimal conditions to guaranty optimal results for the patients.
authors.affiliation: Unité de biologie, Département de biologie et pathologie des tumeurs, Centre GF Leclerc, Dijon, France.
authors.forename: Emeline
authors.initials: E
authors.lastname: Saitta
authors.affiliation: Unité de biologie, Département de biologie et pathologie des tumeurs, Centre GF Leclerc, Dijon, France.
authors.forename: Astrid
authors.initials: A
authors.lastname: Brasselet
authors.affiliation: Unité de biologie, Département de biologie et pathologie des tumeurs, Centre GF Leclerc, Dijon, France.
authors.forename: Muriel
authors.initials: M
authors.lastname: Cadouot
authors.affiliation: Unité de biologie, Département de biologie et pathologie des tumeurs, Centre GF Leclerc, Dijon, France.
authors.forename: Anne
authors.initials: A
authors.lastname: Gibeaud
authors.affiliation: Unité de biologie, Département de biologie et pathologie des tumeurs, Centre GF Leclerc, Dijon, France.
authors.forename: Marine
authors.initials: M
authors.lastname: Jourdain
authors.affiliation: Unité de biologie, Département de biologie et pathologie des tumeurs, Centre GF Leclerc, Dijon, France.
authors.forename: Didier
authors.initials: D
authors.lastname: Bozonnet
authors.affiliation: Unité de biologie, Département de biologie et pathologie des tumeurs, Centre GF Leclerc, Dijon, France.
authors.forename: Vincent
authors.initials: V
authors.lastname: Goussot
authors.affiliation: Unité de biologie, Département de biologie et pathologie des tumeurs, Centre GF Leclerc, Dijon, France.
authors.forename: Jean-Marc
authors.initials: JM
authors.lastname: Riedinger
authors.affiliation: Unité de biologie, Département de biologie et pathologie des tumeurs, Centre GF Leclerc, Dijon, France.
authors.forename: Sarab
authors.initials: S
authors.lastname: Lizard
mesh_terms: D019943:Amino Acid Substitution / Q000235:genetics; D001120:Arginine / Q000235:genetics; D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics; D002478:Cells, Cultured; D004252:DNA Mutational Analysis / Q000379:methods* / Q000592:standards; D066246:ErbB Receptors / Q000235:genetics*; D006801:Humans; D007930:Leucine / Q000235:genetics; D057230:Limit of Detection; D008175:Lung Neoplasms / Q000235:genetics; D061986:MCF-7 Cells; D020125:Mutation, Missense; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D001120:Arginine; C512478:EGFR protein, human; D066246:ErbB Receptors; D007930:Leucine
keywords: EGFR-TKIs; L858R mutation; method validation; non-small lung cancer