Objective: Patients whose cervical cytological exams produced a result of atypical squamous cells of undetermined significance (ASCUS) were asked to undergo human papillomavirus (HPV DNA) genotyping detection to assess the role of HPV infection in ASCUS. Materials and Methods: This study included 1,219 patients with ASCUS that were randomly divided into two groups. The first group contained 618 patients. These participants underwent colposcopy with cervical biopsy. The remaining 601 underwent colposcopy and biopsy with HPV DNA detection. Results: Out of the 56,000 patients with ASCUS who underwent ThinPrep cytology test (TCT) detection in the authors’ hospitals’ gynecological outpatient clinics, 1,604 were diagnosed with ASCUS (2.86%). Among the 1,219 patients with ASCUS, the rate of detection of cervical intraepithelial neoplasia (CIN) and cancerization was 22.89% (279/1,219). Among the 601 patients who underwent HPV testing, 182 were positive for high-risk HPV (30.28%). Among HPV-positive samples, the most common high-risk types were HPV16, and HPV58. The most common low-risk types were HPV6 and HPV11. The rate of detection among highrisk patients who were positive for HPV and cervical carcinoma with intraepithelial neoplasia was 70.88% (129/182). The rate of detection for HPV-negative patients with cervical cancer with intraepithelial neoplasia was 11.55% (47/407). The rate of detection of high-risk HPV was higher than among patients who had not undergone HPV detection and among patients who were negative for HPV (p < 0.05). Conclusion: The results of cervical cytological examination showed that the manner of progression from inflammation to cancer could differ considerably. HPV DNA examination is an effective means of categorizing and managing ASCUS.