The Value of Triple Testing With p16/Ki-67 Dual Staining, Liquid-Based Cytology, and High-Risk HPV in Correlation with Histopathology in Precancerous Cervical Lesions

Objective: To evaluate the diagnostic performance of triple testing—high-risk HPV (HR-HPV) genotyping, liquid-based cytology (LBC), and p16/Ki-67 dual staining—in detecting precancerous cervical lesions, and to assess 2-year follow-up cytology in women without initial biopsy. Methods: A total of 320 women underwent triple testing. Histopathology was available for 232 women and classified as CIN1, CIN2, CIN3, carcinoma in situ (CIS), or adenocarcinoma in situ (AIS). CIN2+ was defined as the primary diagnostic endpoint. Sensitivity, specificity, positive and negative predictive values, and accuracy were calculated for each test. The remaining 88 women without baseline histology entered a prospective follow-up arm and completed repeat cytology after 2 years. Results: Among 232 women with histopathology, 194 (83.6%) had CIN2+ and 38 (16.4%) had CIN1. HR-HPV testing showed the highest sensitivity (96.9%) but low specificity in this referral population. Cytology demonstrated sensitivity of 86.6% and the highest specificity (47.4%). p16/Ki-67 dual staining showed high sensitivity (94.8%) with low specificity (10.5%). Triple-positive profiles (HPV+/abnormal cytology/p16+) strongly correlated with CIN3+. In the follow-up group (n=88), all women were p16/Ki-67–negative at baseline; HPV was positive in 22 (25.0%). After 2 years, cytology results were NILM in 38 (43.2%), CIN1 in 30 (34.1%), and ASC-US in 20 (22.7%), with no CIN2+ cytologic abnormalities observed. Conclusion: Triple testing provides a comprehensive and highly sensitive diagnostic approach for detecting precancerous cervical lesions. The absence of CIN2+ during 2-year follow-up supports the safety of surveillance in HPV-negative and p16/Ki-67–negative women.