Implementation of primary human papillomavirus (HPV) screening will require triage of high-risk (hrHPV)-positive women to efficiently identify those with high risk of cervical high-grade intraepithelial neoplasia (CIN) and cancer, but equally importantly, to deselect hrHPV-positive women who are at low risk . Here, we evaluate the QiaSure methylation assay measuring the human biomarkers FAM19A4 and mir124-2 in combination.
Post-cytology residual SurePath samples from a group of 502 hrHPV positive women undergoing routine cytology screening at Hvidovre Hospital, Denmark, were collected (age 30-65, average: 49 years). HPV testing was done using Onclarity HPV test (BD Diagnostics, Sparks, MD) or CLART2 HPV array (Genomica, Madrid). Women with cytology abnormalities and/or hrHPV positive were referred to follow-up in concordance with Danish Guidelines. In total, 361 of the 502 women had histology registered in the Danish Pathology Databank within 12 months after the positive screening sample. Samples were reflex tested using the QiaSure methylation assay (Qiagen, Hilden, Germany). All molecular testing was performed in concordance with manufacturer’s specification. Clinical performance estimates of reflex methylation for the detection of ≥CIN3 were determined.
Among the 361 women with histology, 8 had CxCa, 54 CIN1, 24 CIN2, 61 CIN3, 214 were normal and 1 had inadequate histology. For ≥CIN3, hrHPV/methylation analysis showed 77% sensitivity, PPV of 40% and NPV of 93%.
In a primary screening setting for women ≥30 years of age, where referral for colposcopy directed biopsies is defined by hrHPV status, the use of QiaSure methylation assay works with SurePath collected cervical samples. The resulting sensitivity, PPV and NPV support that the QiaSure methylation assay can be considered as part of a unified molecular work flow for future molecular cervical screening, saving laboratories the work load of reflex cytology on hrHPV positive screening samples.
This work was supported by the SME Instrument in the Horizon 2020 Work Programme of the European Commission (Valid-screen 666800).