The 2022 global mpox outbreak highlighted the critical importance of public health emergency preparedness. Although mpox is not a new disease, its sudden emergence and rapid spread in many countries, including Cameroon, raised numerous concerns about the ability of health systems to respond effectively. This study aimed to assess the level of preparedness of health districts in the Littoral Region of Cameroon in the event of a potential mpox outbreak, to identify strengths, weaknesses, and priority areas for intervention. We conducted a descriptive cross-sectional study in the health districts of the Littoral Region of Cameroon from August to September 2024. A grid for evaluating preparedness for a potential mpox outbreak was developed and administered to the district health management teams. This tool was based on the Integrated Disease Surveillance and Response (IDSR) technical guidelines. Data collected were analyzed using Microsoft Excel 2016. A total of 17 health districts were surveyed. Regarding their level of preparedness, 5 districts (29%) showed insufficient preparedness (Nkondjock, Loum, Manoka, Ndom, Pouma); 3 districts (17%) had limited preparedness (Boko, Manjo, Dibombari); 7 districts (41%) had an average level of preparedness (Bangue, Edea, Logbaba, Ngambe, Yabassi, Njombe Penja, Nkongsamba); and only 2 districts (11%) demonstrated good preparedness (Newbell, Mbanga). No district in the region reached the optimal level of preparedness. Significant disparities were observed among the districts, with preparedness scores ranging from 0.16 (Nkondjock) to 3.21 (Newbell). Epidemiological surveillance and laboratory capacity had the highest preparedness level (average preparedness score of 3.01), in contrast to case management (insufficient preparedness score of 1.07). The main barriers to preparedness included the absence of a specific mpox response plan, inadequate allocation of financial and material resources, insufficient diagnostic and laboratory confirmation capacity, lack of clinical management protocols, limited training of health personnel on case management, and absence of a risk communication plan. The health districts in the Littoral Region of Cameroon generally exhibited a limited level of preparedness. Several obstacles to preparedness were identified. To prevent a potential mpox outbreak, it is essential to strengthen district-level preparedness, particularly in coordination and response management, case management, risk communication and community engagement, and prevention and control measures, while adopting a differentiated approach that accounts for disparities between districts.