Sensitive detection of FGFR3 mutations in bladder cancer and urine sediments by peptide nucleic acid-mediated real-time PCR clamping

Makito Miyake, Kokichi Sugano, Kiyotaka Kawashima, Hiroki Ichikawa, Kaoru Hirabayashi, Tetsuro Kodama, Hiroyuki Fujimoto, Tadao Kakizoe, Yae Kanai, Kiyohide Fujimoto, Yoshihiko Hirao

研究成果: Article査読

29 被引用数 (Scopus)

抄録

Somatic mutations of the fibroblast growth factor receptor 3 (FGFR3) gene were detected by peptide nucleic acid (PNA)-mediated real-time PCR clamping. Mutation was detected in negative control containing only wild-type DNA due to a misincorporation of dNTPs to PNA binding sites when the amount of template DNA was decreased to 1 ng. Thus, the amount of template DNA was critical determinant of the assay sensitivity in PNA-mediated PCR clamping. Assay conditions were optimized to detect FGFR3 mutations in exons 7, 10, and 15, at a concentration of more than 1% mutated DNA using 50 ng of genomic DNA as the template. Mutations were detected in 12 of 13 (92.3%) tumor tissues and 11 of 13 (84.6%) urine samples from patients with superficial bladder cancer, while no mutations were detected in tissues and/or urine samples from patients with muscle-invasive bladder cancer or chronic cystitis.

本文言語English
ページ(範囲)865-871
ページ数7
ジャーナルBiochemical and Biophysical Research Communications
362
4
DOI
出版ステータスPublished - 2007 11 3
外部発表はい

ASJC Scopus subject areas

  • 生物理学
  • 生化学
  • 分子生物学
  • 細胞生物学

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