AUTHOR=Omic Haris , Vecsei David , Eder Michael , Abd El-Ghany Karim , Winnicki Wolfgang , Schmidt Alice , Kapps Sebastian , Gerges Daniela , Strassl Robert , Wagner Ludwig , Eskandary Farsad TITLE=Urinary VP1 Flow Cytometry as a Complementary Approach for BK Polyomavirus Monitoring: A Proof-Of-Concept Study JOURNAL=Transplant International VOLUME=Volume 39 - 2026 YEAR=2026 URL=https://www.frontierspartnerships.org/journals/transplant-international/articles/10.3389/ti.2026.15780 DOI=10.3389/ti.2026.15780 ISSN=1432-2277 ABSTRACT=Polyomavirus nephropathy (BKPyVAN) is a major cause of allograft dysfunction after kidney transplantation (KTX). While plasma BKPyV-PCR is the diagnostic gold standard, it may not fully reflect tissue injury. We conducted a prospective observational proof-of-concept study in 30 KTX recipients with BKPyV reactivation (November 2022–February 2024); 21 underwent kidney biopsy, 11 were diagnosed with biopsy-proven (BP)-BKPyVAN. Urine samples were analyzed by flow cytometry to quantify the potential of VP1-positive reno-urinary epithelial cells as a novel non-invasive marker of active tubular damage. The control cohort included 21 virology-negative patients. Median urinary VP1-positivity was higher in BP-BKPyVAN (33%, IQR 27–46) vs. non-BKPyVAN patients (5%, IQR 1–13; p < 0.001). The assay achieved an AUC of 0.98 (95% CI 0.93–1.00, p = 0.0003; cut-off: 11.7%; sensitivity = 91%, specificity = 89%) for BP-BKPyVAN. Longitudinally, median VP1-burden declined from 13% (IQR 4–29) at baseline to 0% (IQR 0–0.4). BKPyV-DNAemia declined rapidly, but plateaued at ∼4 × 102–7 × 102 copies/mL, whereas urinary VP1-positive cells became undetectable. Our preliminary results suggest that combining urinary VP1-positivity with plasma BKPyV-PCR may help distinguish BP-BKPyVAN from non-BKPyVAN within a BKPyV-reactivation cohort. Longitudinal VP1 tracking may indicate resolution of viral infection earlier than DNAemia. These findings are hypothesis-generating and require validation in larger independent cohorts.