Clinical Validation of the Venus HPV Full-Genotyping Assay for Cervical Cancer Screening in the VALGENT-4 Framework

Abstract

The Venus HPV assay (VenusHPV) is a real-time PCR-based human papillomavirus (HPV) test that is widely used in China but lacks extensive clinical validation. The VALidation of HPV GENotyping Tests (VALGENT) framework is an established protocol for evaluating HPV genotyping assays against a standard comparator test. This study aimed to assess the clinical accuracy and reproducibility of the VenusHPV assay following international validation criteria. The clinical performance of VenusHPV was evaluated against the GP5+/6+ PCR-based enzyme immunoassay (GP-EIA) using the VALGENT-4 panel, which included 998 consecutive routine screening samples enriched with 297 samples with abnormal cytology from the Danish cervical cancer screening program. Cases were defined as women diagnosed with histologically confirmed cervical intraepithelial neoplasia 2 or more (CIN2+), while two consecutive negative cytology results served as a proxy for nondisease. Intra- and interlaboratory reproducibility was assessed on 500 samples. Using the manufacturer-recommended cutoff, VenusHPV demonstrated noninferior sensitivity for detection of CIN2+, but its specificity for ≤ CIN1 was inferior to that of GP-EIA. Applying an optimized a posteriori cutoff improved the specificity, yielding relative specificity of 1.02 (95% CI [CI], 1.00–1.03; p noninferiority [p_n.inf] < 0.0001), while maintaining a noninferior sensitivity (of 1.02; CI, 1.00–1.08; p_n.inf < 0.0001). The intra- and interlaboratory reproducibility was excellent (95.2%, CI, 93.3–97.1%, Kappa [κ] = 0.87 and 94.0%, CI, 92.0%–96.0%, κ = 0.85, respectively). Notably, the reproducibility criteria were met consistently, regardless of whether the unadjusted or optimized cutoff was applied. The VenusHPV was as sensitive as the GP-EIA for detecting cervical precancer using the unadjusted cutoff but less specific. However, after cutoff optimization, VenusHPV met the international accuracy criteria for cervical cancer screening. Additionally, the assay demonstrated excellent reproducibility.