title: Development of a multiplexed urine assay for prostate cancer diagnosis.
abstract: BACKGROUND
  Several studies have demonstrated the value of DNA methylation in urine-based assays for prostate cancer diagnosis. However, a multicenter validation with a clinical prototype has not been published.
  
  
  METHODS
  We developed a multiplexed, quantitative methylation-specific polymerase chain reaction (MSP) assay consisting of 3 methylation markers, GSTP1, RARB, and APC, and an endogenous control, ACTB, in a closed-tube, homogeneous assay format. We tested this format with urine samples collected after digital rectal examination from 234 patients with prostate-specific antigen (PSA) concentrations > or =2.5 microg/L in 2 independent patient cohorts from 9 clinical sites.
  
  
  RESULTS
  In the first cohort of 121 patients, we demonstrated 55% sensitivity and 80% specificity, with area under the curve (AUC) 0.69. In the second independent cohort of 113 patients, we found a comparable sensitivity of 53% and specificity of 76% (AUC 0.65). In the first cohort, as well as in a combined cohort, the MSP assay in conjunction with total PSA, digital rectal examination status, and age improved the AUC without MSP, although the difference was not statistically significant. Importantly, the GSTP1 cycle threshold value demonstrated a good correlation (R = 0.84) with the number of cores found to contain prostate cancer or premalignant lesions on biopsy. Moreover, samples that exhibited methylation for either GSTP1 or RARB typically contained higher tumor volumes at prostatectomy than those samples that did not exhibit methylation.
  
  
  CONCLUSIONS
  These data confirm and extend previously reported studies and demonstrate the performance of a clinical prototype assay that should aid urologists in identifying men who should undergo biopsy.
authors.affiliation: Veridex LLC, Johnson & Johnson Company, Warren, NJ 07059, USA.
authors.forename: Tatiana
authors.initials: T
authors.lastname: Vener
authors.forename: Carlo
authors.initials: C
authors.lastname: Derecho
authors.forename: Jonathan
authors.initials: J
authors.lastname: Baden
authors.forename: Haiying
authors.initials: H
authors.lastname: Wang
authors.forename: Yashoda
authors.initials: Y
authors.lastname: Rajpurohit
authors.forename: Joanne
authors.initials: J
authors.lastname: Skelton
authors.forename: Jyoti
authors.initials: J
authors.lastname: Mehrotra
authors.forename: Shobha
authors.initials: S
authors.lastname: Varde
authors.forename: Dondapati
authors.initials: D
authors.lastname: Chowdary
authors.forename: Walt
authors.initials: W
authors.lastname: Stallings
authors.forename: Bradley
authors.initials: B
authors.lastname: Leibovich
authors.forename: Howard
authors.initials: H
authors.lastname: Robin
authors.forename: Alexandre
authors.initials: A
authors.lastname: Pelzer
authors.forename: Georg
authors.initials: G
authors.lastname: Schäfer
authors.forename: Marco
authors.initials: M
authors.lastname: Auprich
authors.forename: Sebastian
authors.initials: S
authors.lastname: Mannweiler
authors.forename: Peter
authors.initials: P
authors.lastname: Amersdorfer
authors.forename: Abhijit
authors.initials: A
authors.lastname: Mazumder
mesh_terms: D025601:Adenomatous Polyposis Coli Protein / Q000235:genetics; D000368:Aged; D000369:Aged, 80 and over; D015331:Cohort Studies; D019175:DNA Methylation; D051549:Glutathione S-Transferase pi / Q000235:genetics; D006801:Humans; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods; D011471:Prostatic Neoplasms / Q000175:diagnosis* / Q000473:pathology / Q000652:urine; D018168:Receptors, Retinoic Acid / Q000235:genetics; D012680:Sensitivity and Specificity
chemical_list: D025601:Adenomatous Polyposis Coli Protein; D018168:Receptors, Retinoic Acid; C082925:retinoic acid receptor beta; C496556:GSTP1 protein, human; D051549:Glutathione S-Transferase pi