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High-Sensitivity Detection of Urinary Extracellular Vesicles With Upconverting Nanoparticle-Based Lateral Flow

Md Khirul Islam1,2, Imran Mahmud1,2, Klinton Ali1

  • 1Biotechnology Division, Department of Life Technologies and FICAN West Cancer Centre University of Turku Turku Finland.

Journal of Extracellular Biology
|July 8, 2025
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Summary

A new upconverting nanoparticle lateral flow immunoassay (UCNP-LFIA) detects CD63-positive urinary extracellular vesicles (uEVs). This sensitive test shows significantly higher uEV levels in bladder cancer patients, promising for point-of-care diagnostics.

Keywords:
bladder cancerlateral flow assayrapid testtetraspanin CD63upconversion luminescenceurinary EVs

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Area of Science:

  • Biomedical Diagnostics
  • Nanotechnology in Medicine
  • Urology

Background:

  • Urinary extracellular vesicles (uEVs) are abundant in tetraspanin membrane proteins.
  • CD63 is a key surface marker on uEVs.
  • Current methods for uEV detection can be complex and time-consuming.

Purpose of the Study:

  • To develop a rapid, high-sensitivity upconverting nanoparticle (UCNP)-based lateral flow immunoassay (UCNP-LFIA).
  • To detect CD63-positive uEVs directly from urine for urinalysis.
  • To evaluate the diagnostic potential of UCNP-LFIA for bladder cancer detection.

Main Methods:

  • Developed a sandwich UCNP-LFIA using anti-CD63 antibodies conjugated to UCNPs.
  • Tested minimally processed uEV samples from bladder cancer, benign prostatic hyperplasia, and healthy individuals.
  • Quantified CD63-positive EVs using an upconversion luminescence reader after 80 minutes.

Main Results:

  • The UCNP-LFIA achieved a limit of detection of 4 × 10^7 EVs/mL.
  • CD63-positive EVs were significantly elevated in bladder cancer patients (2.3-fold vs. benign, 16-fold vs. healthy).
  • Demonstrated high sensitivity and quantitative detection of uEVs.

Conclusions:

  • UCNP-LFIA is a sensitive and rapid platform for detecting CD63-positive uEVs.
  • Elevated CD63-positive uEVs show potential as a biomarker for bladder cancer.
  • The UCNP-LFIA holds promise for point-of-care urinalysis and cancer diagnostics.