Lan Xu

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

ABSTRACTThe 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 [pn.inf] < 0.0001), while maintaining a noninferior sensitivity (of 1.02; CI, 1.00–1.08; pn.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.

Evaluation and Optimization of the Clinical Accuracy of Hybribio's 14 High-Risk HPV with 16/18 Genotyping Assay within the VALGENT-3 Framework

Hybribio’s 14 High-Risk HPV with 16/18 genotyping real-time PCR (HBRT-H14) is a human papillomavirus (HPV) assay with approval from the China Food and Drug Administration that is widely used in China. VALGENT (VALidation of HPV GENotyping Tests) is an established framework for evaluating HPV tests’ clinical performance relative to validated comparators. The aim of this study was to assess the clinical accuracy of HBRT-H14 following international validation criteria. Within VALGENT-3, clinical performance of HBRT-H14 was compared with Hybrid Capture 2 (HC2), Linear Array HPV genotyping test (Linear Array), and Cobas 4800 HPV test (Cobas).

Clinical and analytical performance of the CLART HPV 4S assay with SurePath screening samples from the Danish cervical cancer screening program using the VALGENT framework

The CLART HPV4S (CLART4S) is a novel full genotyping assay, based on PCR/microarray technology. We assessed the clinical accuracy of the CLART4S assays under the fourth installment of the VALGENT framework. The VALGENT cohort comprised 998 consecutive cervical samples from women participating in the Danish screening programme enriched with 297 samples with abnormal cytology (100 ASCUS, 100 LSIL, 97 HSIL). The CLART4S assay detects 16 HPV genotypes individually: 14 oncogenic and two non-oncogenic HPV types. The GP5+/6+ PCR Enzyme-Immuno-Assay (GP-EIA) and GP5+/6+ PCR with Luminex genotyping (GP-LMNX) were used as comparator tests for clinical accuracy and HPV genotype concordance, respectively. The sensitivity for ≥ CIN2 for the CLART4S assay was 96.7 % (GP-EIA: 92.6 %) with a relative sensitivity of 1.04 (1.00-1.09). The sensitivity for ≥ CIN3 was 98.8 % (GP-EIA: 94.0 %), with relative sensitivity of 1.05 (1.00-1.10). The specificity for <CIN2 was 88.6 % (GP-EIA: 89.2 %) with a relative specificity of 0.99 (0.98-1.01). The CLART 4S was found to be non-inferior to that of GP-EIA for both sensitivity (p < 0.0001) and specificity (p = 0.0452). The overall oncogenic HPV concordance between CLART4S and GP-LMNX was high, however when looking at individual genotype agreement the concordance was more diverse, with the highest agreement found in the Screening population.

Clinical and Analytical Evaluation of the Alinity m HR HPV Assay within the VALGENT-3 Framework

Only clinically validated human papillomavirus (HPV) tests should be used in cervical cancer screening. VALGENT provides a framework to validate new HPV tests.