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<0.001), with superior sensitivity (74.7% vs 47.0%, P<0.001) and comparable specificity (94.0% vs 94.1%, 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.