title: A multiplex immunoassay for the non-invasive detection of bladder cancer.
abstract: BACKGROUND
  Urine based assays that can non-invasively detect bladder cancer (BCa) have the potential to reduce unnecessary and invasive procedures. The purpose of this study was to develop a multiplex immunoassay that can accurately and simultaneously monitor ten diagnostic urinary protein biomarkers for application as a non-invasive test for BCa detection.
  
  
  METHODS
  A custom electrochemiluminescent multiplex assay was constructed (Meso Scale Diagnostics, LLC, Rockville, MD, USA) to detect the following urinary proteins; IL8, MMP9, MMP10, ANG, APOE, SDC1, A1AT, PAI1, CA9 and VEGFA. Voided urine samples from two cohorts were collected prior to cystoscopy and samples were analyzed blinded to the clinical status of the participants. Means (±SD) and receiver operating characteristic (ROC) curve analysis were used to compare assay performance and to assess the diagnostic accuracy of the diagnostic signature.
  
  
  RESULTS
  Comparative diagnostic performance analyses revealed an AUROC value of 0.9258 for the multiplex assay and 0.9467 for the combination of the single-target ELISA assays (p = 0.625), so there was no loss of diagnostic utility for the MSD multiplex assay. Analysis of the independent 200-sample cohort using the multiplex assay achieved an overall diagnostic sensitivity of 0.85, specificity of 0.81, positive predictive value 0.82 and negative predictive value 0.84.
  
  
  CONCLUSIONS
  It is technically feasible to simultaneously monitor complex urinary diagnostic signatures in a single assay without loss of performance. The described protein-based assay has the potential to be developed for the non-invasive detection of BCa.
authors.affiliation: University of Hawaii Cancer Center, 701 Ilalo St, Rm 327, Honolulu, HI, 96813, USA. yoshikos@hawaii.edu.|Department of Molecular Biosciences and Bioengineering, University of Hawaii at Manoa, Honolulu, HI, USA. yoshikos@hawaii.edu.
authors.forename: Yoshiko
authors.initials: Y
authors.lastname: Shimizu
authors.affiliation: University of Hawaii Cancer Center, 701 Ilalo St, Rm 327, Honolulu, HI, 96813, USA. hfuruya@cc.hawaii.edu.
authors.forename: Hideki
authors.initials: H
authors.lastname: Furuya
authors.affiliation: University of Hawaii Cancer Center, 701 Ilalo St, Rm 327, Honolulu, HI, 96813, USA. pgreenwood@queens.org.
authors.forename: Peter
authors.initials: P
authors.lastname: Bryant Greenwood
authors.affiliation: University of Hawaii Cancer Center, 701 Ilalo St, Rm 327, Honolulu, HI, 96813, USA. ochan@cc.hawaii.edu.
authors.forename: Owen
authors.initials: O
authors.lastname: Chan
authors.affiliation: Department of Biostatistics, University of Florida, Gainesville, FL, USA. ydai@ichp.ufl.edu.
authors.forename: Yunfeng
authors.initials: Y
authors.lastname: Dai
authors.affiliation: Cancer Prevention Program, Fred Hutchinson Cancer Research Center, Seattle, WA, USA. mthornqu@fredhutch.org.
authors.forename: Mark D
authors.initials: MD
authors.lastname: Thornquist
authors.affiliation: Nonagen BioScience Corp, Jacksonville, FL, USA. goodison.steven@mayo.edu.|Department of Health Sciences Research, Mayo Clinic, Jacksonville, FL, USA. goodison.steven@mayo.edu.
authors.forename: Steve
authors.initials: S
authors.lastname: Goodison
authors.affiliation: University of Hawaii Cancer Center, 701 Ilalo St, Rm 327, Honolulu, HI, 96813, USA. cjrosser@hawaii.edu.|Nonagen BioScience Corp, Jacksonville, FL, USA. cjrosser@hawaii.edu.
authors.forename: Charles J
authors.initials: CJ
authors.lastname: Rosser
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D014408:Biomarkers, Tumor / Q000652:urine; D015331:Cohort Studies; D003710:Demography; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D008297:Male; D008875:Middle Aged; D009361:Neoplasm Invasiveness; D015203:Reproducibility of Results; D001749:Urinary Bladder Neoplasms / Q000175:diagnosis* / Q000473:pathology* / Q000652:urine
chemical_list: D014408:Biomarkers, Tumor
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