OBJECTIVE: High-risk human papillomavirus (HR-HPV) persistence is thought to be necessary for the development of cervical cancer. Because not all cases of low-grade squamous intraepithelial lesion (LSIL) or higher-grade squamous intraepithelial lesion are associated with HR-HPV, detection of HR-HPV in women with these cervical abnormalities may be clinically useful for management. The aim of this study was to detect HR-HPV in women with LSIL or higher-grade cytology. MATERIALS AND METHODS: In a multicenter, prospective clinical study, residual liquid-based cytological specimens from 3,966 US women were analyzed. Women with LSIL or higher-grade cytology underwent colposcopy and cervical biopsy as clinically indicated. The test was used to detect HR-HPV in women with cytologically diagnosed LSIL or higher, with histological confirmation of cervical intraepithelial neoplasia 2 (CIN 2) or worse by a central review panel. RESULTS: Among subjects with LSIL or higher-grade cytology and complete data sets, 11.8% (41/347) were diagnosed with CIN 2 or worse; 82.1% (285/347) were HR-HPV positive. The prevalence of CIN 2 or worse and CIN 3 or worse was 14.4% (41/285) and 7.0% (20/285), respectively, among subjects who were HR-HPV positive. All subjects diagnosed with CIN 2 or worse (41/41) and CIN 3 or worse (20/20) tested positive for HR-HPV. Sensitivity and negative predictive value calculated for CIN 2 or worse were 100% (95% CI = 91.4%-100.0%) and 100% (95% CI = 94.2%-100.0%), respectively. Specificity and positive predictive value among these subjects were 20.3% (95% CI = 16.1%-25.1%) and 14.4% (95% CI = 10.8%-18.9%), respectively. CONCLUSIONS: Detection of HR-HPV in women with LSIL or higher-grade cytology using the HPV HR test may allow clinicians to further triage women for advanced clinical management.