BACKGROUND: Cervical screening using primary human papilloma virus (HPV) testing and cytology is being implemented in several countries. Cytology as triage for colposcopy referral suffers from several shortcomings. HPV testing overcomes some of these but lacks specificity in women under 30. Here, we aimed to develop and validate an automatable triage test that is highly sensitive and specific independently of age and sample heterogeneity, and predicts progression to CIN3+ in HPV+ patients. RESULTS: The WID™-qCIN, assessing three regions in human genes DPP6, RALYL, and GSX1, was validated in both a diagnostic (case-control) and predictive setting (nested case-control), in a total of 761 samples. Using a predefined threshold, the sensitivity of the WID™-qCIN test was 100% and 78% to detect invasive cancer and CIN3, respectively. Sensitivity to detect CIN3+ was 65% and 83% for women < and ≥ 30 years of age. The specificity was 90%. Importantly, the WID™-qCIN test identified 52% of ≥ 30-year-old women with a cytology negative (cyt-) index sample who were diagnosed with CIN3 1-4 years after sample donation. CONCLUSION: We identified suitable DNAme regions in an epigenome-wide discovery using HPV+ controls and CIN3+ cases and established the WID™-qCIN, a PCR-based DNAme test. The WID™-qCIN test has a high sensitivity and specificity that may outperform conventional cervical triage tests and can in an objective, cheap, and scalable fashion identify most women with and at risk of (pre-)invasive cervical cancer. However, evaluation was limited to case-control settings and future studies will assess performance and generalisability in a randomised controlled trial.

Cancer Epidemiology Research Programme Catalan Institute of Oncology IDIBELL Av Gran Vía 199 203 08908 L'Hospitalet de Llobregat Barcelona Spain

Center for Health Decision Science Boston MA USA

Consortium for Biomedical Research in Epidemiology and Public Health CIBERESP Carlos 3 Institute of Health Av De Monforte de Lemos 5 28029 Madrid Spain

Department of Epidemiology Harvard T H Chan School of Public Health Boston MA USA

Department of Gynecology and Obstetrics 1st Faculty of Medicine and Hospital Na Bulovce Charles University Prague Prague Czech Republic

Department of Health Policy and Management Harvard T H Chan School of Public Health Boston MA USA

Department of Laboratory Medicine Division of Pathology Karolinska Institutet Stockholm Sweden

Department of Women's and Children's Health Karolinska Institutet Stockholm Sweden

Department of Women's Cancer UCL EGA Institute for Women's Health University College London 74 Huntley Street London WC1E 6AU UK

European Translational Oncology Prevention and Screening Institute Universität Innsbruck Milser Straße 10 6060 Hall in Tirol Austria

Gynaecologic Oncology Center Department of Obstetrics and Gynecology 1st Faculty of Medicine Charles University Prague General University Hospital Prague Prague Czech Republic

Institute for Technology Assessment and Department of Radiology Massachusetts General Hospital Harvard Medical School Boston MA USA

Institute of Public Health Medical Decision Making and Health Technology Assessment Department of Public Health Health Services Research and Health Technology Assessment UMIT TIROL University for Health Sciences and Technology Hall in Tirol Austria

Medical Diagnostics Karolinska Karolinska University Hospital Stockholm Sweden

Research Institute for Biomedical Aging Research Universität Innsbruck 6020 Innsbruck Austria

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