The BD ProEx C stain (BD Diagnostics-Tripath, Burlington, NC) detects 2 molecular markers known to be overexpressed in cervical squamous cell carcinoma, minichromosome maintenance protein 2 and topoisomerase II. This study was conducted to evaluate this stain detecting high-grade cervical disease, and results were compared with high-risk HPV (hr-HPV) status and histologic biopsy diagnosis in Papanicolaou tests with atypical squamous cells, cannot exclude high-grade squamous intraepithelial lesion (ASC-H) cytologic results. Cervical cytology specimens were collected in BD SurePath preservative (BD Diagnostics-Tripath), and 100 specimens with a diagnosis of ASC-H were included in the study. A second BD SurePath slide, prepared from the residual cellular material from these ASC-H cases, was immunostained using the BD ProEx C reagent. Nuclear staining of ASC-H cells was considered a positive result. The sensitivity for BD ProEx C for detecting cervical intraepithelial neoplasia 2 or worse was 98.8% and for hr-HPV was 71.1%, whereas specificity values were 82.4% and 64.7%, respectively. BD ProEx C has potential as a marker for detection of high-grade disease in patients with ASC-H cytologic results.