© 2016 Wiley Periodicals, Inc.Background: Current cervical cancer screening guidelines recommend a 1-year follow-up period for patients with a postmenopausal low-grade squamous intraepithelial lesion (LSIL) who are test negative for high-risk human papillomavirus (HrHPV). The aim of this study was to assess whether such patients had an increased immediate risk of high-grade squamous intraepithelial lesion. Methods: We assessed 54 HrHPV-negative women with postmenopausal LSIL in the Department of Obstetrics and Gynecology of our hospital between 2012 and 2013. All patients underwent liquid-based cytology and reflex HrHPV testing (for human papillomavirus [HPV] types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68). Colposcopic examination and guided biopsy were performed by the same gynecologist (MO). Results: The average age of the patients was 53.1 ± 3.2 years. There were 33 patients (61%) with cervical intraepithelial neoplasia (CIN) grade 1 and 21 who were non-dysplastic. None of the patients was positive for CIN 2 or any other lesions. Conclusions: If the HPV test is negative, repeat cytology after 12 months is recommended by the American Society for Colposcopy and Cervical Pathology for cases of HrHPV-negative postmenopausal LSIL. We recommend reflex HPV testing as the best choice for patients who test positive for postmenopausal LSIL by Pap smear, in line with the literature. Diagn. Cytopathol. 2016;44:969–974. © 2016 Wiley Periodicals, Inc.