title: Comparative analysis of prostate cancer specific biomarkers PCA3 and ERG in whole urine, urinary sediments and exosomes.
abstract: BACKGROUND
  PCA3 and ERG are mRNA-based prostate cancer (PCa) specific biomarkers that can be detected in urine. However, urine is a complex substrate that can be separated in several fractions. In this study we compared the levels of PCa-specific biomarkers (PCA3 and ERG) and KLK3 as prostate-specific reference gene in three urine substrates-whole urine, urinary sediment (cell pellet) and exosomes-and evaluated the influence of performing a digital rectal examination (DRE) prior to urine sampling.
  
  
  METHODS
  First-voided urine samples were prospectively obtained before and after DRE from 29 men undergoing prostate biopsies. The urine was separated in whole urine, cell pellet and exosomes and the biomarker levels were measured with RT-qPCR.
  
  
  RESULTS
  PCa was identified in 52% (15/29) of men. In several samples the mRNA levels were below the analytical limit of detection (BDL). The biomarker levels were highest in whole urine and significantly higher after DRE in all substrates. In PCa patients higher levels of PCA3 and ERG were found in all urine substrates after DRE compared to non-PCa patients.
  
  
  CONCLUSIONS
  This is the first study in which urinary PCa-specific biomarker levels were compared directly in three separate urine fractions. These results suggest that whole urine could be the urine substrate of choice for PCa-diagnostics based on analytical sensitivity, which is reflected directly in the high informative rate. Moreover, the significant positive effect of performing a DRE prior to urine sampling is confirmed. These findings could be of influence in the development of PCa-diagnostic urine tests.
authors.forename: Rianne J
authors.initials: RJ
authors.lastname: Hendriks
authors.forename: Siebren
authors.initials: S
authors.lastname: Dijkstra
authors.forename: Sander A
authors.initials: SA
authors.lastname: Jannink
authors.forename: Martijn G
authors.initials: MG
authors.lastname: Steffens
authors.forename: Inge M
authors.initials: IM
authors.lastname: van Oort
authors.forename: Peter F A
authors.initials: PF
authors.lastname: Mulders
authors.forename: Jack A
authors.initials: JA
authors.lastname: Schalken
mesh_terms: D000368:Aged; D000951:Antigens, Neoplasm / Q000235:genetics / Q000652:urine*; D014408:Biomarkers, Tumor / Q000235:genetics / Q000652:urine*; D055354:Exosomes / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D008297:Male; D008875:Middle Aged; D011471:Prostatic Neoplasms / Q000652:urine*; D012333:RNA, Messenger / Q000652:urine; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000071230:Transcriptional Regulator ERG / Q000235:genetics / Q000652:urine
chemical_list: D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; C072312:ERG protein, human; D012333:RNA, Messenger; D000071230:Transcriptional Regulator ERG; C479341:prostate cancer antigen 3, human