Abstract 4917: A four-gene methylation marker panel as triage test in hr-HPV positive patients

Objective: Currently no cervical neoplasia specific methylation markers with high sensitivity and specificity are available for use in population-based screening on (pre)malignant cervical neoplasia. Aim of the study was to identify new methylation markers and to design and validate a methylation marker panel for triage of hr-HPV positive patients. Methods: Quantitative methylation specific PCRs (MSP) on OpenArray™ platform, representing 424 primers of 213 cancer specific methylated genes, were performed on frozen tissue samples from 84 cervical cancer patients and 106 normal cervices. The top 20 of ranked methylation markers were validated by LightCycler® MSP experiments. Then the top 3 methylation markers based on ROC analysis were selected for further clinical validation in combination with C13ORF18 (previously identified by our group), on cervical scrapings from 74 cervical cancer patients, 69 normal cervices and 148 patients referred with an abnormal Pap smear. Finally, a scenario analysis for population based screening program was performed to compare the diagnostic performance of our methylation panel to conventional liquid based cytology after primary hr-HPV testing. Results: Three cervical neoplasia specific methylation markers (JAM3, EPB41L3 and TERT) discriminated strongly between cervical scrapings from cervical cancer patients and healthy controls (p