TY - JOUR
T1 - Sensitive detection of FGFR3 mutations in bladder cancer and urine sediments by peptide nucleic acid-mediated real-time PCR clamping
AU - Miyake, Makito
AU - Sugano, Kokichi
AU - Kawashima, Kiyotaka
AU - Ichikawa, Hiroki
AU - Hirabayashi, Kaoru
AU - Kodama, Tetsuro
AU - Fujimoto, Hiroyuki
AU - Kakizoe, Tadao
AU - Kanai, Yae
AU - Fujimoto, Kiyohide
AU - Hirao, Yoshihiko
PY - 2007/11/3
Y1 - 2007/11/3
N2 - 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.
AB - 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.
KW - FGFR3
KW - Mutation
KW - Peptide nucleic acid
KW - Real-time PCR
KW - Superficial bladder cancer
UR - http://www.scopus.com/inward/record.url?scp=34548583630&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=34548583630&partnerID=8YFLogxK
U2 - 10.1016/j.bbrc.2007.08.092
DO - 10.1016/j.bbrc.2007.08.092
M3 - Article
C2 - 17803960
AN - SCOPUS:34548583630
VL - 362
SP - 865
EP - 871
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 4
ER -