Introduction: Patient transport employees frequently come into contact with multidrug-resistant organisms (MDROs) and therefore are at a greater risk of infection than the general population. These pathogens pose a significant challenge for employees of patient transport services since they can spread over long distances through patient transfers. To date, little is known about the occupational risk of MRSA infection in patient transport settings. Methods: A cross-sectional study was conducted to investigate the prevalence of MRSA in patient transport personnel, including taxi drivers, as well as the potential risk factors for MRSA colonization. For screening, nasal swabs were taken. When an individual was tested positive, a control swab was taken; if this confirmed a positive result, decolonization measures were offered. A molecular biological examination of the MRSA samples was performed. Results: A total of 222 patient transport employees were screened and 7 employees tested positive, putting the MRSA prevalence at 3.2% (95% CI 1.4-6.5). Significant risk factors among patient transport staff (PTS) for testing positive were the use of antibiotics (OR 11.9; 95% CI 1.8-78.4) and hospital admission (OR 6.9; 95% CI 1.1-45.9). MRSA swabs were also performed on a total of 102 taxi drivers who provide patient transport services. The MRSA prevalence was 0.98 (95% CI <0.01-5.9). Significant group differences between PTS and taxi drivers, with respect to potential risk factors for MRSA colonization, were identified as inpatient treatment (p=0.09), chronic respiratory illnesses (p=0.01), and knowingly transporting patients/passengers with MRSA (p=0.03). Conclusion: This study is the first to make data on the MRSA risk of patient transport employees in Hamburg available. The prevalence data are low in all areas and indicate a somewhat low risk of infection. A good infection control at the facilities is highly recommendable and the employees should acquire in-depth knowledge of infection prevention to improve compliance with personal protective measures.