title: Rapid detection of epidermal growth factor receptor mutations in lung cancer by the SMart-Amplification Process.
abstract: OBJECTIVE
  A positive response to gefitinib in non-small cell lung cancer (NSCLC) has been correlated to mutations in epidermal growth factor receptor (EGFR) gene. Previous reports have been based mainly on diagnostic screening by sequencing. However, sequencing is a time-consuming and complicated procedure, not suitable for routine clinical use.
  
  
  METHODS
  We have developed rapid, simple, and sensitive mutation detection assays based on the SMart Amplification Process (SMAP) and applied it for analyzing EGFR gene mutations in clinical samples. By using SMAP, we can detect mutations within 30 min including sample preparation. To validate the assay system for potential use in clinical diagnostics, we examined 45 NSCLC patients for EGFR mutations using sequencing and SMAP.
  
  
  RESULTS
  The outcomes of the SMAP assay perfectly matched the sequencing results, except in one case where SMAP was able to identify a mutation that was not detected by sequencing. We also evaluated the sensitivity and specificity of SMAP in mutation detection for EGFR. In a serial dilution study, SMAP was able to find a mutation in a sample containing only 0.1% of the mutant allele in a mixture of wild-type genomic DNA. We also could show amplification of mutated DNA with only 30 copies per reaction.
  
  
  CONCLUSIONS
  The SMAP method offers higher sensitivity and specificity than alternative technologies, while eliminating the need for sequencing to identify mutations in the EGFR gene of NSCLC. It provides a robust and point-of-care accessible approach for a rapid identification of most patients likely to respond to gefitinib.
authors.affiliation: Genome Exploration Research Group (Genome Network Project Core Group), RIKEN Genomic Sciences Center, RIKEN Yokohama Institute, 1-7-22 Suehiro-cho, Japan. Canaty@aol.com
authors.forename: Kanako
authors.initials: K
authors.lastname: Hoshi
authors.forename: Hideki
authors.initials: H
authors.lastname: Takakura
authors.forename: Yasumasa
authors.initials: Y
authors.lastname: Mitani
authors.forename: Kenji
authors.initials: K
authors.lastname: Tatsumi
authors.forename: Nobuyoshi
authors.initials: N
authors.lastname: Momiyama
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Ichikawa
authors.forename: Shinji
authors.initials: S
authors.lastname: Togo
authors.forename: Toru
authors.initials: T
authors.lastname: Miyagi
authors.forename: Yuki
authors.initials: Y
authors.lastname: Kawai
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Kogo
authors.forename: Takeshi
authors.initials: T
authors.lastname: Kikuchi
authors.forename: Chiaki
authors.initials: C
authors.lastname: Kato
authors.forename: Takahiro
authors.initials: T
authors.lastname: Arakawa
authors.forename: Syuji
authors.initials: S
authors.lastname: Uno
authors.forename: Paul E
authors.initials: PE
authors.lastname: Cizdziel
authors.forename: Alexander
authors.initials: A
authors.lastname: Lezhava
authors.forename: Noburou
authors.initials: N
authors.lastname: Ogawa
authors.forename: Yoshihide
authors.initials: Y
authors.lastname: Hayashizaki
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Shimada
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics*; D045744:Cell Line, Tumor; D066246:ErbB Receptors / Q000235:genetics*; D005091:Exons; D005260:Female; D017353:Gene Deletion; D005838:Genotype; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D009154:Mutation; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012680:Sensitivity and Specificity