Abstract
BACKGROUND: Cervical cancer remains a significant global health concern, necessitating effective screening strategies. Traditional methods, such as liquid-based cytology (LBC) and high-risk human papillomavirus (HR-HPV) testing, have limitations in sensitivity and specificity. This study aimed to evaluate the diagnostic performance of p16, Ki-67, and minichromosome maintenance protein 2 (MCM2) as biomarkers in cervical cancer screening, both as standalone methods and in combination, to improve early detection and risk stratification. METHODS: This prospective study included 344 women who underwent LBC and HR-HPV testing, followed by the gold standard of colposcopy and biopsy. Immunocytochemical (ICC) staining for p16, Ki-67, and MCM2 was performed. The sensitivity, specificity, and Youden index were calculated to compare the efficacy of the single and combined screening methods. RESULTS: LBC alone demonstrated suboptimal sensitivity, while HR-HPV testing exhibited low specificity. Among the single methods, dual staining of p16 paired with MCM2 (p16/MCM2) when using a high-risk threshold achieved the highest Youden index (0.55). The combination of LBC with p16/MCM2 yielded superior sensitivity (96.3%) and moderate specificity (42.6%), outperforming the combination of HR-HPV with dual staining of p16/MCM2. Importantly, all missed diagnoses were high-grade squamous intraepithelial lesion (HSIL) with cervical intraepithelial neoplasia (CIN) grades 2-3, and no cases of cervical cancer were missed. CONCLUSIONS: HR-HPV-based primary screening with p16/Ki-67 triage may offer a viable strategy for cervical cancer detection. Combining LBC with p16/MCM2 dual staining demonstrated improved sensitivity and specificity compared to conventional methods. Notably, p16/MCM2 as a standalone assay achieved the highest Youden index when using a high-risk threshold. Further validation through multicenter studies is essential to confirm its generalizability.