biostudies-literatureBell MBill and Melinda Gates Foundation (GF)World Health OrganizationEuropean Research CouncilUniversity of Antwerp (DOCPRO4-TT(ZAP)BOF)Fonds Wetenschappelijk Onderzoek (FWO)e0140424https://www.ebi.ac.uk/biostudies/studies/S-EPMC11837537biostudies-literatureUnknown63(2)Despite first-void urine (FVU) being increasingly recognized as a credible specimen for human papillomavirus (HPV) detection, there is a lack of well-validated testing methods providing full quantitative genotyping required for vaccine impact monitoring from FVU samples. The Allplex HPV28 assay, capable of individually detecting 28 HPV genotypes, presents a promising method. We aimed to evaluate its genotype-specific performance on FVU samples, following optimization of FVU preanalytics. We selected 701 FVU samples collected using a Colli-Pee device (20 mL, with UCM), enriched for HPV-positivity (<i>n</i> = 630) based on previous testing with GP5+/6+-PCR-based reverse line blot (GP5+/6+ RLB) and E7-MPG after Amicon filtration (AF). We first evaluated the comparability and agreement of AllpJournal of clinical microbiologyOptimization and analytical validation of the Allplex HPV28 genotyping assay for use in first-void urine samples.PMC1183753735537001OPP1053353101040588FN54230000511PJK24NTenet VHeideman DAMGheit TBell MClifford GVan Caesbroeck ARol MVorsters ABaussano IfalseOptimization and analytical validation of the Allplex HPV28 genotyping assay for use in first-void urine samples.Despite first-void urine (FVU) being increasingly recognized as a credible specimen for human papillomavirus (HPV) detection, there is a lack of well-validated testing methods providing full quantitative genotyping required for vaccine impact monitoring from FVU samples. The Allplex HPV28 assay, capable of individually detecting 28 HPV genotypes, presents a promising method. We aimed to evaluate its genotype-specific performance on FVU samples, following optimization of FVU preanalytics. We selected 701 FVU samples collected using a Colli-Pee device (20 mL, with UCM), enriched for HPV-positivity (<i>n</i> = 630) based on previous testing with GP5+/6+-PCR-based reverse line blot (GP5+/6+ RLB) and E7-MPG after Amicon filtration (AF). We first evaluated the comparability and agreement of Allp2025-01-01T00:00:00Z2025 Feb2025-04-04T02:05:11.298Z2025-04-04T02:05:11.298ZS-EPMC118375373972383610.1128/jcm.01404-24