Reproducibility of HPV Testing With Two Versions of the Papilloplex High‐Risk HPV Assay

ABSTRACT

Validation of HPV tests usable in cervical cancer screening require demonstration of noninferior clinical accuracy and sufficient reproducibility. The Papilloplex HR‐HPV assay [version1] (Genefirst, UK) has been validated only for clinical accuracy. Here, we assessed its reproducibility, aiming to complete the validation process. A panel of 550 cryopreserved cervical cell samples collected from women attending the cervical cancer screening program in Belgium was used to assess the intra‐ and inter‐laboratory reproducibility of Papilloplex HR‐HPV [version1], a full genotyping assay that identifies separately 14 high‐risk HPV (hrHPV) types using multiple probe amplification technology. We assessed whether the reproducibility fulfils validation criteria (lower 95% confidence interval [CI] bound ≥ 87% and κ ≥ 0.50). Subsequently, we compared the concordance between version1 and a new version6 of the assay and between three analysis methods for PCR curve interpretation. Papilloplex HR‐HPV version1 assay showed an excellent reproducibility for hrHPV (97.5% [CI: 95.8%–98.7%], κ = 0.94 for intra‐ and 93.5% [CI: 91.0%–95.4%], κ = 0.85 for inter‐laboratory reproducibility). Concordance analyses exhibited an excellent agreement between two assay versions and between three PCR curve analysis methods. Papilloplex HR‐HPV version1 assay exhibited excellent reproducibility, completing the international validation criteria. Papilloplex HR‐HPV version6 showed excellent concordance with version1 but still lacks clinical validation.