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

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.