title: A 3-plex methylation assay combined with the FGFR3 mutation assay sensitively detects recurrent bladder cancer in voided urine.
abstract: OBJECTIVE
  DNA methylation is associated with bladder cancer and these modifications could serve as useful biomarkers. FGFR3 mutations are present in 60% to 70% of non-muscle invasive bladder cancer (NMIBC). Low-grade bladder cancer recurs in more than 50% of patients. The aim of this study is to determine the sensitivity and specificity of a urine assay for the diagnosis of recurrences in patients with a previous primary NMIBC G1/G2 by using cystoscopy as the reference standard.
  
  
  METHODS
  We selected eight CpG islands (CGI) methylated in bladder cancer from our earlier genome-wide study. Sensitivity of the CGIs for recurrences detection was investigated on a test set of 101 preTUR urines. Specificity was determined on 70 urines from healthy males aged more than 50 years. A 3-plex assay for the best combination was developed and validated on an independent set of 95 preTUR, recurrence free, and nonmalignant urines (n=130).
  
  
  RESULTS
  The 3-plex assay identified recurrent bladder cancer in voided urine with a sensitivity of 74% in the validation set. In combination with the FGFR3 mutation assay, a sensitivity of 79% was reached (specificity of 77%). Sensitivity of FGFR3 and cytology was 52% and 57%, respectively.
  
  
  CONCLUSIONS
  The combination of methylation and FGFR3 assays efficiently detects recurrent bladder cancer without the need for stratification of patients regarding methylation/mutation status of the primary tumor. We conclude that the sensitivity of this combination is in the same range as cystoscopy and paves the way for a subsequent study that investigates a modified surveillance protocol consisting of the urine test followed by cystoscopy only when the urine test is positive.
authors.affiliation: Department of Pathology, Erasmus MC, Rotterdam, the Netherlands.
authors.forename: Raju
authors.initials: R
authors.lastname: Kandimalla
authors.forename: Roy
authors.initials: R
authors.lastname: Masius
authors.forename: Willemien
authors.initials: W
authors.lastname: Beukers
authors.forename: Chris H
authors.initials: CH
authors.lastname: Bangma
authors.forename: Torben F
authors.initials: TF
authors.lastname: Orntoft
authors.forename: Lars
authors.initials: L
authors.lastname: Dyrskjot
authors.forename: Nikki
authors.initials: N
authors.lastname: van Leeuwen
authors.forename: Hester
authors.initials: H
authors.lastname: Lingsma
authors.forename: Angela A G
authors.initials: AA
authors.lastname: van Tilborg
authors.forename: Ellen C
authors.initials: EC
authors.lastname: Zwarthoff
mesh_terms: D000368:Aged; D014408:Biomarkers, Tumor / Q000235:genetics* / Q000652:urine; D018899:CpG Islands / Q000235:genetics; D019175:DNA Methylation / Q000235:genetics; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D009154:Mutation; D009364:Neoplasm Recurrence, Local / Q000235:genetics* / Q000473:pathology / Q000652:urine; D009367:Neoplasm Staging; D051498:Receptor, Fibroblast Growth Factor, Type 3 / Q000235:genetics* / Q000652:urine; D001749:Urinary Bladder Neoplasms / Q000235:genetics* / Q000473:pathology / Q000652:urine
chemical_list: D014408:Biomarkers, Tumor; C496365:FGFR3 protein, human; D051498:Receptor, Fibroblast Growth Factor, Type 3