Abstract

Background: Persistent infection with high-risk Human Papillomavirus (HR-HPV) is the main cause of cervical cancer. However, current screening methods remain limited in distinguishing transient infections from clinically significant lesions requiring further intervention. p16/Ki-67 dual staining is an immunocytochemical assay that detects cellular alterations associated with HPV-mediated oncogenic transformation and has been increasingly incorporated into cervical cancer screening programs. This study aimed to provide a preliminary assessment of the feasibility of p16/Ki-67 dual staining on liquid-based cervical cytology samples at the University Medical Center Ho Chi Minh City. Objective: To standardize and preliminarily assess the implementation of p16/Ki-67 dual staining (CINtec® PLUS Cytology) on liquid-based cervical cytology samples at the University Medical Center Ho Chi Minh City. Materials and Methods: The study included HR-HPV-positive cases with liquid-based cervical cytology (LiquiPrep) at the University Medical Center Ho Chi Minh City. Method verification followed CAP 2024 and CLSI EP12-IG 2023 guidelines, and feasibility was assessed in 39 cases (January - April 2026). Dual staining was performed using the CINtec® PLUS kit on the Ventana BenchMark ULTRA system. Slides were evaluated for staining quality, interpretability, technical artifacts, and correlation with cytology and histopathology results. Results: The method met verification criteria. Among 25 cases with histopathological correlation, all CIN2+ and malignant lesions were positive while most lesions ≤CIN1 were negative. Observed technical artifacts did not significantly affect interpretation. Conclusion: p16/Ki-67 dual staining is feasible on liquid-based cervical cytology samples, providing adequate slide quality, good interpretability, and acceptable technical artifacts, with results consistent with histopathological severity.