title: Clinical screening assay for EGFR exon 19 mutations using PNA-clamp smart amplification process version 2 in lung adenocarcinoma.
abstract: The presence of EGFR mutations is correlated with a positive therapeutic response to tyrosine kinase inhibitors; therefore, the accurate detection of EGFR mutations is crucial when deciding appropriate therapeutic strategies. Recently, the rapid and sensitive assay smart amplification process version 2 (SmartAmp2) was developed. However, this method can only detect one type of mutation in EGFR exon 19; therefore, we applied the PNA technology to the SmartAmp2 assay to develop PNA-clamp SmartAmp2 for the detection of many types of deletions in EGFR exon 19, in a single reaction. This new assay was evaluated using 172 clinical samples. Thirty-nine (22.7%) samples were found to have deletions by PNA-clamp SmartAmp2; whereas 30 (17.4%) and 38 (22.1%) tumors were found to have deletions by direct sequencing and PNA-enriched sequencing, respectively. Three cases, in which we detected mutations with PNA-clamp SmartAmp2, but not with direct sequencing, were treated with gefitinib, and all cases showed a partial therapeutic response. Using clinical samples, we demonstrated that PNA-clamp SmartAmp2 can detect various types of mutations in EGFR exon 19 in a relatively short time and with high sensitivity. This method detected small amounts of mutant DNA and identified patients for whom clinical information was previously unavailable from other tests. This test may contribute to the administration of efficient therapeutic strategies.
authors.affiliation: Department of Clinical Pharmacology, Gunma University Graduate School of Medicine, 3-39-22 Showa-machi, Maebashi 371-8511, Japan. tkyaraki@gunma-u.ac.jp
authors.forename: Takuya
authors.initials: T
authors.lastname: Araki
authors.forename: Kimihiro
authors.initials: K
authors.lastname: Shimizu
authors.forename: Tomonori
authors.initials: T
authors.lastname: Nakamura
authors.forename: Masaru
authors.initials: M
authors.lastname: Baba
authors.forename: Yuki
authors.initials: Y
authors.lastname: Kawai
authors.forename: Katsunori
authors.initials: K
authors.lastname: Nakamura
authors.forename: Yasumasa
authors.initials: Y
authors.lastname: Mitani
authors.forename: Kyoko
authors.initials: K
authors.lastname: Obayashi
authors.forename: Tohru
authors.initials: T
authors.lastname: Aomori
authors.forename: Yukiyoshi
authors.initials: Y
authors.lastname: Fujita
authors.forename: Yohei
authors.initials: Y
authors.lastname: Miyamae
authors.forename: Seiichi
authors.initials: S
authors.lastname: Kakegawa
authors.forename: Kyoichi
authors.initials: K
authors.lastname: Kaira
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 / Q000201:enzymology / Q000235:genetics*; D000077192:Adenocarcinoma of Lung; D001483:Base Sequence; D045744:Cell Line, Tumor; D004252:DNA Mutational Analysis / Q000379:methods; D066246:ErbB Receptors / Q000235:genetics*; D005091:Exons; D006801:Humans; D008175:Lung Neoplasms / Q000201:enzymology / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D020135:Peptide Nucleic Acids / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods
chemical_list: D020135:Peptide Nucleic Acids; D066246:ErbB Receptors