Clinical Validation of Four Point-of-Care High-Risk HPV Assays, Including Two Reduced-Valency Assays, for Cervical...

Persistent infection with high-risk human papillomavirus (HR-HPV) is the necessary cause of cervical cancer, with approximately 95% of cases attributable to eight most carcinogenic HPV types (16/18/31/33/35/45/52/58). Several HPV assays have recently been developed in India for use in cervical cancer screening, including reduced-valency tests targeting seven/eight oncogenic HPV types. We evaluated the clinical accuracy and reproducibility of four such assays: PathoDetect-HPV-14, HPV-Q (14 types), Truenat-HR-HPV-Plus (8 types), and PathoDetect-HPV-7 (7 types). Using the VALGENT framework, 1159 cervical samples from the ESTAMPA study conducted in Argentina and Costa Rica were analysed with the new assays. The samples included 97 cases of CIN2+ (19 CIN2, 72 CIN3, and 6 cancers). Assay performance was compared in a blinded manner with established reference tests (Cobas-4800 and Allplex-HPV-HR) and reduced-valency comparators. Sensitivity, specificity, agreement, and repeatability were assessed. Truenat-HR-HPV-Plus demonstrated a sensitivity of 80.4% (95% CI: 71.1-87.8) and specificity of 91.5% (95% CI: 89.5-93.2) for CIN2+. It met IARC validation criteria and showed non-inferior performance to Allplex-HPV-HR-8, with relative sensitivity of 1.03 (95% CI: 0.96-1.09) for CIN2+ and 1.00 (95% CI: 0.96-1.04) for CIN3+, and relative specificity of 0.99 (95% CI: 0.97-1.00) and 0.98 (95% CI: 0.97-1.00), respectively. Repeatability was 93.3% (κ = 0.79). PathoDetect-HPV-7 showed lower sensitivity (68.1%) with specificity of 89.0%. HPV-Q and PathoDetect-HPV-14 did not meet validation criteria. This study represents the first formal validation of reduced-valency HPV assays and demonstrates that Truenat-HR-HPV-Plus provides robust clinical performance with higher specificity than 14-valent assays, supporting its potential to improve screening efficiency and reduce unnecessary referrals.