A Novel Multiplex ELISA Assay for Evaluating Patients With Gross Hematuria for Bladder Cancer

Hematuria is the most common presentation of BCa with 22% of patients with gross hematuria harboring BCa. VUC is the most widely used urine-based assay for detecting BCa; however, it fails to detect approximately 50% of low-grade or early stage BCa when it is most curable.

Because of this severe limitation, patients with hematuria (microscopic or gross) will undergo an invasive examination of the urinary bladder, where a miniature camera is inserted into the bladder. We propose to improve upon the non-invasive detection of BCa by further validating a multiplex ELISA assay directed at a BCa-associated diagnostic signature in voided urine samples of patients with gross hematuria.

Primary outcomes

• To examine the sensitivity and specificity of the multiplex ELISA assay directed at a BCa-associated diagnostic signature in voided urine samples to reference standard of cystoscopy in a cohort of 450 participants presenting with gross hematuria. [1 year]

Secondary outcomes

• To compare the sensitivity and specificity of the multiplex ELISA assay to VUC and NMP22® BladderChek to reference standard of cystoscopy in this cohort. [1 year]
• To develop a BCa risk calculator from this cohort using cutting edge machine learning techniques (e.g., random forest) incorporating biomarker data and clinical data. [1 year]

Sex: All, ages: 18 Years to any

Inclusion Criteria:

* Age 18 years or older
* Have documented or reported gross hematuria within 3 month of study enrollment
* Willing and able to give written informed consent

Exclusion Criteria (patients must not):

* Have history of BCa
* History of previous cancer (excluding basal and squamous cell skin cancer) within the past 3 years
* Have a known active urinary tract infection or urinary retention
* Have active stone disease (renal or bladder) or renal insufficiency (creatinine >2.0 mg/dL) Serum creatinine value can be up to 60 days before consent, otherwise repeat
* Have ureteral stents, nephrostomy tubes or bowel interposition
* Have recent genitourinary instrumentation (within 10 days prior to signing consent)
* Be unable or unwilling to complete the hematuria evaluation (i.e., cystoscopy and upper tract imaging)

• Results publication: Lotan Y, Gupta A, Miyake M, Anai S, Luu M, Ahdoot M, Messing E, Peers G, Chin AI, Liu M, Tikhonekov S, Sakatani T, Shimizu T, Pagano I, Zheng Y, Zhang Z, Kim H, Josephson D, Furuya H, Rosser CJ. Prospective multicenter validation of a Urine-Based multiplex assay for risk stratification of patients presenting with gross hematuria. J Natl Cancer Inst. 2026 Aug 17:djag282. doi: 10.1093/jnci/djag282. Online ahead of print. PMID 42608795
• Results publication: Furuya H, Sakatani T, Tanaka S, Murakami K, Waldron RT, Hogrefe W, Rosser CJ. Bladder cancer risk stratification with the Oncuria 10-plex bead-based urinalysis assay using three different Luminex xMAP instrumentation platforms. J Transl Med. 2024 Jan 2;22(1):8. doi: 10.1186/s12967-023-04811-2. PMID 38167321
• Results publication: Furuya H, Sakatani T, Tanaka S, Murakami K, Waldron RT, Hogrefe W, Rosser CJ. Bladder cancer risk stratification with the Oncuria 10-plex bead-based urinalysis assay using three different Luminex xMAP instrumentation platforms. Res Sq [Preprint]. 2023 Nov 25:rs.3.rs-3635581. doi: 10.21203/rs.3.rs-3635581/v1. PMID 38045238

• Informed Consent Form (PDF, 2024-05-31)

• Charles Rosser, MD, Study director, Nonagen Bioscience Corporation
• Hideki Furuya, MD, Principal investigator, Cedars-Sinai Medical Center

NIH grants: R01CA198887