title: EGFR and KRAS mutational analysis in a large series of Italian non-small cell lung cancer patients: 2,387 cases from a single center.
abstract: Activating EGFR mutations are important genetic alterations that have strong therapeutic implications for non-small cell lung cancer (NSCLC) patients. However, the role of KRAS mutations in this process is still under evaluation. Here, we report on the feasibility of a large‑scale EGFR and KRAS mutation analysis in the daily routine of a single center. NSCLCs from 2,387 patients were screened for EGFR and KRAS mutations from January 2010 to September 2015. Mutational analyses were performed in a single laboratory using single strand conformation polymorphism (SSCP)-Sanger sequencing and matrix‑assisted laser desorption ionization‑time of flight (MALDI‑TOF) on Sequenom platform for EGFR and pyrosequencing for KRAS. Activating EGFR mutations were found in 14.1% of all tumors, whereas KRAS mutations were found in 30.5% of all tumors. Direct sequencing showed analyzable cytological, small biopsy and surgical specimen percentages of 90.3, 90.9 and 98.1%, respectively, whereas the MALDI‑TOF platform showed analyzable cytological samples, small biopsies and surgical specimens percentages of 94.6, 95.7 and 96.9%, respectively. The mean analytical turnaround times (TAT) were 4 and 3 days for direct sequencing and the MALDI‑TOF platform, respectively. Our results confirm that small biopsy or cytological samples can be used for reliable EGFR and KRAS mutation testing and indicate that adopting the MALDI‑TOF platform reduces the rate of missed samples among the samples. Moreover, the 3-day analytical TAT of the MALDI-TOF multi-target technique is appropriate for clinical management and reduces the overall treatment decision time.
authors.affiliation: Department of Surgical, Medical, Molecular Pathology and Critical Area, University of Pisa, Pisa, Italy.
authors.forename: Riccardo
authors.initials: R
authors.lastname: Giannini
authors.affiliation: Unit of Anatomic Pathology 3, Azienda Ospedaliero‑Universitaria Pisana, Pisa, Italy.
authors.forename: Cristiana
authors.initials: C
authors.lastname: Lupi
authors.affiliation: Unit of Anatomic Pathology 3, Azienda Ospedaliero‑Universitaria Pisana, Pisa, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Sensi
authors.affiliation: Unit of Anatomic Pathology 3, Azienda Ospedaliero‑Universitaria Pisana, Pisa, Italy.
authors.forename: Greta
authors.initials: G
authors.lastname: Alì
authors.affiliation: Unit of Anatomic Pathology 3, Azienda Ospedaliero‑Universitaria Pisana, Pisa, Italy.
authors.forename: Agnese
authors.initials: A
authors.lastname: Proietti
authors.affiliation: Department of Surgical, Medical, Molecular Pathology and Critical Area, University of Pisa, Pisa, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Boldrini
authors.affiliation: Department of Surgical, Medical, Molecular Pathology and Critical Area, University of Pisa, Pisa, Italy.
authors.forename: Adele
authors.initials: A
authors.lastname: Servadio
authors.affiliation: Department of Surgical, Medical, Molecular Pathology and Critical Area, University of Pisa, Pisa, Italy.
authors.forename: Mirella
authors.initials: M
authors.lastname: Giordano
authors.affiliation: Department of Surgical, Medical, Molecular Pathology and Critical Area, University of Pisa, Pisa, Italy.
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Macerola
authors.affiliation: Department of Surgical, Medical, Molecular Pathology and Critical Area, University of Pisa, Pisa, Italy.
authors.forename: Rossella
authors.initials: R
authors.lastname: Bruno
authors.affiliation: Department of Surgical, Medical, Molecular Pathology and Critical Area, University of Pisa, Pisa, Italy.
authors.forename: Nicla
authors.initials: N
authors.lastname: Borrelli
authors.affiliation: Unit of Pneumology, Azienda Ospedaliero‑Universitaria Pisana, Pisa, Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: Chella
authors.affiliation: Unit of Thoracic Surgery, Azienda Ospedaliero‑Universitaria Pisana, Pisa, Italy.
authors.forename: Franca
authors.initials: F
authors.lastname: Melfi
authors.affiliation: Department of Surgical, Medical, Molecular Pathology and Critical Area, University of Pisa, Pisa, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Lucchi
authors.affiliation: Unit of Thoracic Endoscopy, Azienda Ospedaliero‑Universitaria Pisana, Pisa, Italy.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Ribechini
authors.affiliation: Unit of Medical Oncology 2, Azienda Ospedaliero‑Universitaria Pisana, Pisa, Italy.
authors.forename: Enrico
authors.initials: E
authors.lastname: Vasile
authors.affiliation: Medical Oncology Department, Istituto Toscano Tumori, Ospedale Civile, Livorno, Italy.
authors.forename: Federico
authors.initials: F
authors.lastname: Cappuzzo
authors.affiliation: Department of Surgical, Medical, Molecular Pathology and Critical Area, University of Pisa, Pisa, Italy.
authors.forename: Alfredo
authors.initials: A
authors.lastname: Mussi
authors.affiliation: Department of Surgical, Medical, Molecular Pathology and Critical Area, University of Pisa, Pisa, Italy.
authors.forename: Gabriella
authors.initials: G
authors.lastname: Fontanini
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D044963:Biopsy, Fine-Needle / Q000379:methods; D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics*; D004252:DNA Mutational Analysis / Q000379:methods; D066246:ErbB Receptors / Q000235:genetics*; D005260:Female; D006801:Humans; D007558:Italy; D008175:Lung Neoplasms / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D009154:Mutation / Q000235:genetics*; D016283:Proto-Oncogene Proteins p21(ras) / Q000235:genetics*
chemical_list: C117307:KRAS protein, human; C512478:EGFR protein, human; D066246:ErbB Receptors; D016283:Proto-Oncogene Proteins p21(ras)