BACKGROUND: Atypical squamous cells of undetermined significance (ASC-US) is the most frequent but ambiguous abnormal Papanicolaou (Pap) interpretation and is generally triaged by high-risk human papillomavirus (hrHPV) testing before colposcopy. This study aimed to evaluate the performance of an artificial intelligence (AI)-based triage system to predict ASC-US cytology for cervical intraepithelial neoplasia 2+ lesions (CIN2+). METHODS: More than 60,000 images were used to train this proposed deep learning-based ASC-US triage system, where both cell-level and slide-level information were extracted. In total, 1967 consecutive ASC-US Paps from 2017 to 2019 were included in this study. Histological follow-ups were retrieved to compare the triage performance between the AI system and hrHPV in 622 patients with simultaneous hrHPV testing. RESULTS: In the triage of women with ASC-US cytology for CIN2+, our system attained equivalent sensitivity (92.9%; 95% confidence interval [CI], 75.0%-98.8%) and higher specificity (49.7%; 95% CI, 45.6%-53.8%) than hrHPV testing (sensitivity: 89.3%; 95% CI, 70.6%-97.2%; specificity: 34.3%; 95% CI, 30.6%-38.3%) without requiring additional patient examination or testing. Additionally, the independence of this system from hrHPV testing (κ = 0.138) indicated that these 2 different methods could be used to triage ASC-US as an alternative way. CONCLUSION: This de novo deep learning-based system can triage ASC-US cytology for CIN2+ with a performance superior to hrHPV testing and without incurring additional expenses.