A multiplex immunoassay for the non-invasive detection of bladder cancer

Yoshiko Shimizu, Hideki Furuya, Peter Bryant Greenwood, Owen Chan, Yunfeng Dai, Mark D. Thornquist, Steve Goodison, Charles J. Rosser

Research output: Contribution to journalArticlepeer-review

25 Scopus citations


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.

Original languageEnglish (US)
Article number31
JournalJournal of Translational Medicine
Issue number1
StatePublished - Jan 30 2016


  • Biomarkers
  • Bladder cancer
  • Multiplex
  • Protein
  • Urine

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)


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