Analytical and clinical validation of the EdenDx methylation assay as a noninvasive supportive triage tool for early detection of endometrial cancer in cervical scraping specimens

Objective Endometrial cancer (EC) is the most common gynecologic malignancy; however, early detection remains challenging due to overlapping symptoms and the absence of validated noninvasive triage testing. Diagnosis relies on invasive procedures such as endometrial biopsy or dilation and curettage. The EdenDx assay detects hypermethylation of CDO1 and CELF4 genes, specifically associated with EC and have been shown to be epigenetically altered prior to symptom onset. Here, we evaluate the clinical and analytical performance of the EdenDx assay as a noninvasive triage tool for the early detection of endometrial cancer in cervical cytology specimens. Methods Analytical validation including limit of detection (LoD), precision, analytical specificity, adequacy, was performed using contrived specimens. Clinical performance was evaluated in two specimen types: cervical scraping specimens collected from 18 patients with histologically confirmed EC and 62 patients without EC, and formalin-fixed paraffin-embedded (FFPE) tissue from 20 patients with EC and 20 without EC, with histopathology as the reference standard. Results The LoD was 1.5% cancer cells; the coefficient of variation (CV) was <10% across runs and operators, and no interference from common vaginal pathogens or substances was observed. In cervical scraping specimens, sensitivity was 72.2% (13/18; 95% confidence interval [CI], 46.5–90.3%) and specificity was 96.8% (60/62; 95% CI, 88.8–99.6%); in FFPE tissue, sensitivity was 85.0% (17/20) and specificity was 90.0% (18/20); in contrived specimens, both sensitivity and specificity were 100%. Overall, the assay demonstrated pooled sensitivity of 90.2% and specificity of 96.2%. Conclusions The EdenDx assay is analytically robust and detects CDO1/CELF4 hypermethylation in both cervical scraping specimens and FFPE endometrial tissue, with high specificity and sensitivity in cervical scrapings. These findings support further prospective evaluation of the EdenDx assay as a noninvasive triage adjunct to identify high-risk women with or without abnormal uterine bleeding who warrant endometrial sampling.