title: Sensitive detection of FGFR3 mutations in bladder cancer and urine sediments by peptide nucleic acid-mediated real-time PCR clamping.
abstract: 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.
authors.affiliation: Oncogene Research Unit/Cancer Prevention Unit, Tochigi Cancer Center Research Institute, 4-9-13 Yonan, Utsunomiya, Tochigi 320-0834, Japan.
authors.forename: Makito
authors.initials: M
authors.lastname: Miyake
authors.forename: Kokichi
authors.initials: K
authors.lastname: Sugano
authors.forename: Kiyotaka
authors.initials: K
authors.lastname: Kawashima
authors.forename: Hiroki
authors.initials: H
authors.lastname: Ichikawa
authors.forename: Kaoru
authors.initials: K
authors.lastname: Hirabayashi
authors.forename: Tetsuro
authors.initials: T
authors.lastname: Kodama
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Fujimoto
authors.forename: Tadao
authors.initials: T
authors.lastname: Kakizoe
authors.forename: Yae
authors.initials: Y
authors.lastname: Kanai
authors.forename: Kiyohide
authors.initials: K
authors.lastname: Fujimoto
authors.forename: Yoshihiko
authors.initials: Y
authors.lastname: Hirao
mesh_terms: D004252:DNA Mutational Analysis / Q000379:methods*; D015342:DNA Probes / Q000235:genetics; D006801:Humans; D020135:Peptide Nucleic Acids / Q000235:genetics*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D051498:Receptor, Fibroblast Growth Factor, Type 3 / Q000235:genetics* / Q000652:urine*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D016482:Urinalysis / Q000379:methods*; D001749:Urinary Bladder Neoplasms / Q000235:genetics*