Comparative Diagnostic Performance of DNA Ploidy Analysis and p16/Ki67 Dual-Staining with Cytology in hrHPV-Defined Subgroups: A Single-Center Evaluation

Objective: To systematically evaluate the agreement of deoxyribonucleic acid (DNA) ploidy analysis and p16/Ki67 dual‑staining (DS) with cytology, and compare their diagnostic performance for detecting cervical abnormalities, particularly in high-risk human papillomavirus (hrHPV)‑defined subgroups. Methods: This cross-sectional study enrolled 877 women undergoing cervical cancer screening. Cervical exfoliated cells were collected for liquid-based cytology, hrHPV testing, DNA ploidy analysis, and p16/Ki67 dual-staining. Agreement between tests was assessed using kappa statistics. Diagnostic performance for detecting cytological abnormalities (≥ASC-US, ≥LSIL) and histologically confirmed CIN2+ was evaluated using area under the ROC curve (AUC), sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV). Results: DNA ploidy demonstrated stronger agreement with cytology (κ=0.60, 95% CI: 0.51– 0.69) than DS (κ=0.40, 95% CI: 0.30– 0.51). For detecting ≥ASC-US, DNA ploidy achieved significantly higher AUC than DS (0.843 vs 0.705, P 0.999). DNA ploidy showed sensitivity comparable to hrHPV testing (74.7% vs 79.5%) but with significantly higher specificity (94.0% vs 84.4%, P < 0.001). In hrHPV-positive women, DNA ploidy exhibited the strongest agreement with cytology (κ=0.69) and maintained robust diagnostic performance. For CIN2+ detection, DNA ploidy showed sensitivity of 69.2% and specificity of 62.7%, while DS demonstrated lower sensitivity (53.8%). Conclusion: DNA ploidy analysis demonstrates stronger agreement with cytology and superior diagnostic performance compared to p16/Ki67 dual-staining, particularly in hrHPV-positive subgroups where effective triage is essential. With sensitivity comparable to hrHPV testing but significantly higher specificity, DNA ploidy offers a balanced, automated approach maintaining high detection rates in cervical cancer screening programs. Keywords: DNA ploidy, p16/Ki67 dual-staining, cervical cancer