Background: The prevalence of multidrug-resistant organisms (MDROs) in acute healthcare settings is increasing worldwide. Active screening for MDROs carriers on admission permits the prompt implementation of the appropriate precautions to decrease the probability of cross transmission to other inpatients. Objective: To report the spectrum of bacterial nasal, axilla, and perianal colonization among in patients at Najah National University Hospital (NNUH) during 2018. Methods: A retrospective observational study was performed at NNUH, a tertiary-care referral university hospital in Nablus, north of Palestine, that includes medical and surgical ICUs for both adults and children from January to August 2018. Nasal, axilla, and perianal swabs were collected within the first 24 hours of admission according to hospital policy. Patients who were referred from another hospital, who were admitted to a hospital for at least 2 nights during the previous 8 months, and who are known to have an MDROs in the past were included. Swab samples were processed for isolation and identification of these multidrug-resistant strains. Transmission-based precautions were implemented if positive results were reported (ie, contact isolation) and decolonization regimens were applied according to the CDC recommendations (muporocin ointment for nasal MRSA, daily bathing with chlorhexidine 2% soap for the rest). A daily isolation list was circulated among bed managers and senior nurses and head of departments for appropriate management of beds and reallocation of patients. The antibiotic susceptibility pattern was assessed using the disc-diffusion method on Mueller–Hinton agar and a Vitek-2 system. Results: During the period of the study, 1,425 nasal swabs, 1,245 axilla swabs, and 300 perianal swabs were collected according to the inclusion criteria. Positive results were reportedin 7%, 4%, and 44% for nasal, axilla, and perianal specimens, respectively. Regarding the distribution of bacterial colonization in the nasal swab, 73% were MRSA; for the axial, 29% were Pseudomonas; and from the perianal swab, the most prevalent pathogen was ESBL (56%) (Figs. 1–3). A discrepancy between the number of nasal or axilla and perianal swabs was observed, which was mainly due to the refusal of many patients to have the sample collected by the nurse. Conclusions: Colonization of the skin and mucous membranes of inpatients with MDROs is considered a risk factor for developing future infections. Therefore, active screening for those pathogens is critical for infection prevention and control programs and patient safety in acute-care settings.