Atypical squamous cells of undetermined significance (ASCUS) are the most common cytological abnormality of all smear test. No study has demonstrated the prevalence of cervical cancer or its precursor in Chinese patients with ASCUS. This study aims to investigate the prevalence of cervical intraepithelial neoplasia 1 or worse (CIN1+) and CIN3 or worse (CIN3+) in patients with ASCUS in China to provide insight into appropriate management for Chinese health care.In a retrospective cross-sectional study, patients who underwent liquid-based thin layer cytology and human papillomavirus (HPV) co-testing at the Peking Union Medical College Hospital between January 2014 and January 2017, and had ASCUS results on liquid-based thin layer cytology test and underwent follow-up and colposcopic biopsy were included. Age, HPV DNA test, and pathological outcomes were assessed.One hundred forty-four patients with ASCUS and positive HPV test results were included. In the 3-year follow-up, 23 (16.0%) patients had CIN1, 28 (19.4%) had CIN2, and 17 (11.8%) had CIN3 or carcinoma in situ. The risk of CIN3+ was significantly higher in those older than 60 years (42.8%, P = .005), whereas the CIN1+ prevalence displayed no significant difference between age groups. Both hybrid Capture II (HC II) value and cytopathological description of HPV infection showed no statistically significant correlation with CIN1+ or CIN3+.Patients with HPV-positive ASCUS who were older than 60 years had a significantly higher risk of CIN3+, and clinicians should pay more attention to them. Both HC II value and cytopathological description of HPV infection showed no significant correlation with CIN1+ or CIN3+.