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, Steven Goodison, Charles J. Rosser

Research output: Contribution to journalArticle

18 Citations (Scopus)

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.

Original languageEnglish (US)
Article number31
JournalJournal of Translational Medicine
Volume14
Issue number1
DOIs
StatePublished - Jan 30 2016

Fingerprint

Immunoassay
Urinary Bladder Neoplasms
Assays
Urine
Unnecessary Procedures
Cystoscopy
Proteins
Apolipoproteins E
Interleukin-8
ROC Curve
Biomarkers
Enzyme-Linked Immunosorbent Assay
Sensitivity and Specificity

Keywords

  • Biomarkers
  • Bladder cancer
  • Multiplex
  • Protein
  • Urine

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Shimizu, Y., Furuya, H., Bryant Greenwood, P., Chan, O., Dai, Y., Thornquist, M. D., ... Rosser, C. J. (2016). A multiplex immunoassay for the non-invasive detection of bladder cancer. Journal of Translational Medicine, 14(1), [31]. https://doi.org/10.1186/s12967-016-0783-2

A multiplex immunoassay for the non-invasive detection of bladder cancer. / Shimizu, Yoshiko; Furuya, Hideki; Bryant Greenwood, Peter; Chan, Owen; Dai, Yunfeng; Thornquist, Mark D.; Goodison, Steven; Rosser, Charles J.

In: Journal of Translational Medicine, Vol. 14, No. 1, 31, 30.01.2016.

Research output: Contribution to journalArticle

Shimizu, Y, Furuya, H, Bryant Greenwood, P, Chan, O, Dai, Y, Thornquist, MD, Goodison, S & Rosser, CJ 2016, 'A multiplex immunoassay for the non-invasive detection of bladder cancer', Journal of Translational Medicine, vol. 14, no. 1, 31. https://doi.org/10.1186/s12967-016-0783-2
Shimizu Y, Furuya H, Bryant Greenwood P, Chan O, Dai Y, Thornquist MD et al. A multiplex immunoassay for the non-invasive detection of bladder cancer. Journal of Translational Medicine. 2016 Jan 30;14(1). 31. https://doi.org/10.1186/s12967-016-0783-2
Shimizu, Yoshiko ; Furuya, Hideki ; Bryant Greenwood, Peter ; Chan, Owen ; Dai, Yunfeng ; Thornquist, Mark D. ; Goodison, Steven ; Rosser, Charles J. / A multiplex immunoassay for the non-invasive detection of bladder cancer. In: Journal of Translational Medicine. 2016 ; Vol. 14, No. 1.
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AB - 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.

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