Objectives: This study aimed to investigate the prevalence of Low Back Pain (LBP) among female nursing staff and explore the potential risk factors associated with LBP. Methods: An analytical cross-sectional study was conducted on randomly selected female nurses using payroll as a sampling frame in all public hospitals in Sana'a City, Yemen. Data was collected through face-to-face interview using a structured, pre-coded questionnaire that was available in Arabic and English. Weight and height of the nurses were measured using weight and height scales and body mass index was calculated. Multiple logistic regression was used to identify the factors associated with LBP. Results: Out of 696 female nurses selected, 687 (98.7%) responded. The life-time, the 12-month and one-week prevalence rates of LBP among female nurses were 512 (74.5%; 95% CI: 71.1-77.7%), 411 (59.8%; 95% CI: 56.0-63.5%) and 249 (36.2%; 95% CI: 32.6-39.9%), respectively. The prevalence was significantly lower in Indian nurses compared to other nurses. Three out of every 10 nurses with LBP had sick leave because of LBP in the last 12 months. Factors that showed significant association with LBP among nursing staff in the multivariate analysis were age, nationality, menstrual disorders and stress level at work. Conclusion: LBP is common among female nurses in Yemen. The role of menstrual disorders in developing LBP among female nurses seems to be important. Although sharing the same working conditions, Indian nurses were less likely to report LBP, which highlight the importance of cultural differences in willingness to report LBP.