A Simple and Fast Method for the Simultaneous Detection of Nine Fibroblast Growth Factor Receptor 3 Mutations in Bladder Cancer and Voided Urine

• Published in: Clinical cancer research 2005-11, Vol.11 (21), p.7743-7748
• Main Authors: VAN OERS, Johanna M. M, LURKIN, Irene, VAN EXSEL, Antonius J. A, NIJSEN, Yvette, VAN RHIJN, Bas W. G, VAN DER AA, Madelon N. M, ZWARTHOFF, Ellen C
• Format: Article
• Language: English
• Subjects: Antineoplastic agents; Biological and medical sciences; bladder cancer; Deoxyribonucleotides - genetics; DNA - metabolism; DNA Mutational Analysis; DNA Primers - genetics; Electrophoresis, Capillary; FGFR3; Humans; Medical sciences; Mutation; mutation analysis; Nephrology. Urinary tract diseases; Pharmacology. Drug treatments; Polymerase Chain Reaction; Polymorphism, Single-Stranded Conformational; Prognosis; Receptor, Fibroblast Growth Factor, Type 3 - genetics; Sensitivity and Specificity; Sequence Analysis, DNA; Tumors of the urinary system; Urinary Bladder Neoplasms - genetics; Urinary Bladder Neoplasms - urine; Urinary system involvement in other diseases. Miscellaneous; Urinary tract. Prostate gland; urine
• ISSN: 1078-0432; 1557-3265
• DOI: 10.1158/1078-0432.CCR-05-1045

Purpose: Mutations in the fibroblast growth factor receptor 3 ( FGFR3 ) occur in 50% of primary bladder tumors. An FGFR3 mutation is associated with good prognosis, illustrated by significantly lower percentage of patients with progression and disease-specific mortality. FGFR3 mutations are especially prevalent in low grade/stage tumors, with pTa tumors harboring mutations in 85% of the cases. These tumors recur in 70% of patients. Efficient FGFR3 mutation detection for prognostic purposes and for detection of recurrences in urine is an important clinical issue. In this paper, we describe a simple assay for the simultaneous detection of nine different FGFR3 mutations. Experimental Design: The assay consists of one multiplex PCR, followed by extension of primers for each mutation with a labeled dideoxynucleotide. The extended primers are separated by capillary electrophoresis, and the identity of the incorporated nucleotide indicates the presence or absence of a mutation. Results: The assay was found to be more sensitive than single-strand conformation polymorphism analysis. Mutations could still be detected with an input of only 1 ng of genomic DNA and in a 20-fold excess of wild-type DNA. Moreover, in urine samples from patients with a mutant tumor, the sensitivity of mutation detection was 62%. Conclusions: We have developed a fast, easy to use assay for the simultaneous detection of FGFR3 mutations, which can be of assistance in clinical decision-making and as an alternative for the follow-up of patients by invasive cystoscopy for the detection of recurrences in urine.