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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.

journal_name

Biochem Biophys Res Commun

journal_title

Biochemical and biophysical research communications

authors

Miyake M,Sugano K,Kawashima K,Ichikawa H,Hirabayashi K,Kodama T,Fujimoto H,Kakizoe T,Kanai Y,Fujimoto K,Hirao Y

doi

10.1016/j.bbrc.2007.08.092

subject

Has Abstract

pub_date

2007-11-03 00:00:00

pages

865-71

issue

4

eissn

0006-291X

issn

1090-2104

pii

S0006-291X(07)01769-X

journal_volume

362

pub_type

杂志文章

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