DNA methylation markers for the triage of high-risk human papillomavirus infected women

Aim: To identify and validate highly discriminating DNA methylation markers for the triage of hrHPV-positive women with or without clinically relevant lesions.

Methods: Hypermethylated DNA enriched from cervical cancers was compared with that from cervical scrapes of HPV16-positive cases with no evidence for disease by CpG island microarray hybridization. The most promising marker regions were validated by quantitative methylation-specific PCR (qMSP) using DNA from archived cervical tissues and cervical scrapes. The performance of these markers was then determined in an independent set of 217 hrHPV-positive cervical scrapes from outpatients with histopathological verification.

Results: A methylation signature comprising the 5'regions of the genes DLX1, ITGA4, RXFP3, SOX17 and ZNF671 specific for CIN3 and cervical cancer (termed CIN3+) was identified and validated. A high detection rate of CIN3+ was obtained if at least 2 of the 5 markers were methylated. In the subsequent cross-sectional study our marker panel achieved a sensitivity and specificity for CIN3+ of 96.2% (95% CI 80.4 – 100%) and 76.6% (95% CI 65.6 – 85.5%), respectively for women ≥30 years of age. By projecting the observed test performance to the target population of a HPV screening study (n = 3292) we conducted in Eastern Thuringia some years ago, the sensitivity, specificity, PPV and NPV were calculated to be 90.5%, 81.0%, 61.6% and 96.2%, respectively.

Conclusions: Molecular triage of hrHPV-positive women by our methylation panel is feasible and opens the way to full molecular screening.