title: A fully integrated, automated and rapid detection system for KRAS mutations.
abstract: KRAS mutations are detected in tumors of various organs, and they are also markers of resistance for epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors and monoclonal antibodies against the EGFR. Thus, the accurate and rapid detection of KRAS mutations is crucial, not only for screening, but also for the prediction of the efficacy of molecular-targeted therapy. The aim of the present study was to establish a novel automated detection system for KRAS mutations. One hundred and thirty-six lung adenocarcinoma patients were genotyped for KRAS mutations with both the conventional direct sequence (DS) method and with the newly developed quenching probe (QP) method that obtains data automatically within 60 min. The detection limit of the QP method using a control plasmid containing the KRAS mutation was 50 copies, and 10% mutant plasmid was detected in the mixture of wild-type and mutants. The results obtained by the QP and DS methods were identical in all but two of the 136 cases. The two differentially identified samples, which consisted of substantially fewer lung cancer cells, were positive according to the QP method but negative as determined by DS for KRAS mutations. These findings characterize the QP method as an accurate and rapid detection system for KRAS mutations.
authors.affiliation: Division of Hematology, Respiratory Medicine and Oncology, Department of Internal Medicine, Faculty of Medicine, Saga University, and Department of Medical Oncology, Saga Prefectural Hospital, Saga 849-8501, Japan.
authors.forename: Norio
authors.initials: N
authors.lastname: Ureshino
authors.forename: Naoko
authors.initials: N
authors.lastname: Sueoka-Aragane
authors.forename: Tomomi
authors.initials: T
authors.lastname: Nakamura
authors.forename: Akemi
authors.initials: A
authors.lastname: Sato
authors.forename: Kazutoshi
authors.initials: K
authors.lastname: Komiya
authors.forename: Kentaro
authors.initials: K
authors.lastname: Iwanaga
authors.forename: Masahiro
authors.initials: M
authors.lastname: Mitsuoka
authors.forename: Yuji
authors.initials: Y
authors.lastname: Takeda
authors.forename: Shinichiro
authors.initials: S
authors.lastname: Hayashi
authors.forename: Eisaburo
authors.initials: E
authors.lastname: Sueoka
authors.forename: Shinya
authors.initials: S
authors.lastname: Kimura
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics* / Q000473:pathology; D000077192:Adenocarcinoma of Lung; D057205:Automation, Laboratory; D003062:Codon; D004252:DNA Mutational Analysis / Q000379:methods*; D015342:DNA Probes / Q000737:chemistry; D006801:Humans; D057230:Limit of Detection; D008175:Lung Neoplasms / Q000235:genetics* / Q000473:pathology; D009154:Mutation; D009693:Nucleic Acid Hybridization / Q000379:methods; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D016283:Proto-Oncogene Proteins p21(ras); D012235:Rhodamines / Q000737:chemistry; D018631:ras Proteins / Q000235:genetics*
chemical_list: C114557:5-carboxytetramethylrhodamine succinimidyl ester; D003062:Codon; D015342:DNA Probes; C117307:KRAS protein, human; D011518:Proto-Oncogene Proteins; D012235:Rhodamines; D016283:Proto-Oncogene Proteins p21(ras); D018631:ras Proteins