title: Prospective study of the accuracy of EGFR mutational analysis by high-resolution melting analysis in small samples obtained from patients with non-small cell lung cancer.
abstract: OBJECTIVE
  Epidermal growth factor receptor (EGFR) mutations, especially in-frame deletions in exon 19 (DEL) and a point mutation in exon 21 (L858R), predict gefitinib sensitivity in patients with non-small cell lung cancer (NSCLC). In this study, we verified the accuracy of EGFR mutation analysis in small samples by high-resolution melting analysis (HRMA), which is a rapid method using PCR amplification with a dye to analyze the melting curves in NSCLC.
  
  
  METHODS
  We designed a prospective study to compare the sensitivity and specificity of HRMA and DNA sequencing with laser capture microdissection. Eligible patients with lung lesions were screened by bronchoscopy or percutaneous needle biopsy to histologically confirm the diagnosis, followed by surgical resection of the NSCLC. Small diagnostic specimens were analyzed for EGFR mutations by HRMA, and the surgically resected specimens were examined for mutations by HRMA and DNA sequencing.
  
  
  RESULTS
  The analyses for EGFR mutations were conducted in 52 eligible cases of the 92 enrolled patients. EGFR mutations were detected in 18 (34.6%) patients. The results of HRMA from surgically resected specimens as well as DNA sequencing revealed 100% sensitivity and specificity. On the other hand, the sensitivity and specificity of HRMA from the small diagnostic specimens were 83.3% and 100%, respectively.
  
  
  CONCLUSIONS
  In this study, we showed that HRMA is a highly accurate method for detecting DEL and L858R mutations in patients with NSCLC, although it is necessary to consider the identification of patients with a false-negative result when the analysis is conducted using small samples.
authors.affiliation: Division of Internal Medicine, Teikyo University School of Medicine, Tokyo, Japan.
authors.forename: Tomoya
authors.initials: T
authors.lastname: Fukui
authors.forename: Yuichiro
authors.initials: Y
authors.lastname: Ohe
authors.forename: Koji
authors.initials: K
authors.lastname: Tsuta
authors.forename: Koh
authors.initials: K
authors.lastname: Furuta
authors.forename: Hiromi
authors.initials: H
authors.lastname: Sakamoto
authors.forename: Toshimi
authors.initials: T
authors.lastname: Takano
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Nokihara
authors.forename: Noboru
authors.initials: N
authors.lastname: Yamamoto
authors.forename: Ikuo
authors.initials: I
authors.lastname: Sekine
authors.forename: Hideo
authors.initials: H
authors.lastname: Kunitoh
authors.forename: Hisao
authors.initials: H
authors.lastname: Asamura
authors.forename: Takaaki
authors.initials: T
authors.lastname: Tsuchida
authors.forename: Masahiro
authors.initials: M
authors.lastname: Kaneko
authors.forename: Masahiko
authors.initials: M
authors.lastname: Kusumoto
authors.forename: Seiichiro
authors.initials: S
authors.lastname: Yamamoto
authors.forename: Teruhiko
authors.initials: T
authors.lastname: Yoshida
authors.forename: Tomohide
authors.initials: T
authors.lastname: Tamura
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics* / Q000378:metabolism; D004252:DNA Mutational Analysis / Q000379:methods*; D066246:ErbB Receptors / Q000235:genetics* / Q000378:metabolism*; D005260:Female; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics* / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D009154:Mutation; D011237:Predictive Value of Tests; D011446:Prospective Studies; D012680:Sensitivity and Specificity
chemical_list: D066246:ErbB Receptors