title: Usefulness of peptide nucleic acid (PNA)-clamp smart amplification process version 2 (SmartAmp2) for clinical diagnosis of KRAS codon 12 mutations in lung adenocarcinoma: comparison of PNA-clamp SmartAmp2 and PCR-related methods.
abstract: KRAS is an oncogene that can be activated by mutations. Patients with non-small cell lung cancer who have KRAS mutations do not respond to tyrosine kinase inhibitors; therefore, accurate detection of KRAS mutations is important for deciding therapeutic strategies. Although sequencing-related techniques have been frequently used, they are usually too complex, have low sensitivity, and are time-consuming for routine screening in clinical situations. We evaluated peptide nucleic acid (PNA)-clamp smart amplification process version 2 (SmartAmp2) as a detection method for KRAS codon 12 mutations in patient specimens compared with traditional sequencing and polymerase chain reaction-related methods. Among 172 lung adenocarcinoma samples, direct sequencing, enzyme-enriched sequencing, and PNA-enriched sequencing showed that 16 (9.3%), 26 (15.7%), and 28 (16.3%) tumors, respectively, contained KRAS mutations in codon 12. Using PNA-clamp SmartAmp2, we could identify 31 (18.0%) tumors that had KRAS mutations in codon 12 within 60 minutes, three of which were undetected by polymerase chain reaction-related methods. On the other hand, we examined 30 nonmalignant peripheral lung tissue specimens and found no mutations in any of the samples using PNA-clamp SmartAmp2. In this study, we confirmed that PNA-clamp SmartAmp2 has high sensitivity and accuracy and is suitable for the clinical diagnosis of KRAS codon 12 mutations.
authors.affiliation: Department of Clinical Pharmacology, Gunma University Graduate School of Medicine, 3-39-22 Showa-machi, Maebashi 371-8511, Japan.
authors.forename: Takuya
authors.initials: T
authors.lastname: Araki
authors.forename: Kimihiro
authors.initials: K
authors.lastname: Shimizu
authors.forename: Katsunori
authors.initials: K
authors.lastname: Nakamura
authors.forename: Tomonori
authors.initials: T
authors.lastname: Nakamura
authors.forename: Yasumasa
authors.initials: Y
authors.lastname: Mitani
authors.forename: Kyoko
authors.initials: K
authors.lastname: Obayashi
authors.forename: Yukiyoshi
authors.initials: Y
authors.lastname: Fujita
authors.forename: Seiichi
authors.initials: S
authors.lastname: Kakegawa
authors.forename: Yohei
authors.initials: Y
authors.lastname: Miyamae
authors.forename: Kyoichi
authors.initials: K
authors.lastname: Kaira
authors.forename: Takefumi
authors.initials: T
authors.lastname: Ishidao
authors.forename: Alexander
authors.initials: A
authors.lastname: Lezhava
authors.forename: Yoshihide
authors.initials: Y
authors.lastname: Hayashizaki
authors.forename: Izumi
authors.initials: I
authors.lastname: Takeyoshi
authors.forename: Koujirou
authors.initials: K
authors.lastname: Yamamoto
mesh_terms: D000230:Adenocarcinoma / Q000175:diagnosis / Q000235:genetics*; D045744:Cell Line, Tumor; D004252:DNA Mutational Analysis / Q000191:economics / Q000379:methods*; D006801:Humans; D008175:Lung Neoplasms / Q000175:diagnosis / Q000235:genetics*; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000191:economics / Q000379:methods*; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D016283:Proto-Oncogene Proteins p21(ras); D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013997:Time Factors; D018631:ras Proteins / Q000235:genetics*
chemical_list: C117307:KRAS protein, human; D011518:Proto-Oncogene Proteins; D016283:Proto-Oncogene Proteins p21(ras); D018631:ras Proteins