ABSTRACT   BACKGROUND፡ Hospital infections in pediatric units increase the length of hospital stay and the use of antibiotics, and this causes exposure to more procedures. This study was aimed to determine the microorganisms represented in the hand flora of pediatric patients at the beginning of hospitalization and before discharge. METHODS: The study was designed as a prospective cross-sectional study. This prospective study was performed with 124 pediatric patients. After completion of the admission procedures, an initial sample was taken from the hands of the hospitalized patients. Another sample was taken from the patients just before discharging. RESULTS: Growth of coagulase-negative staphylococcus (CNS) was observed in the culture samples of 28 patients. Cultures from 23 patients showed different microorganisms such as Staphylococcus aureus, Escherichia coli, and S. epidermis. Examination of final discharge cultures showed CNS in 43 patients, S. aureus in 5 patients, E. coli in 8 patients, Acinetobacter baumannii in 11 patients, and Kocuria rhizophila, K. kristinae, Candida spp., Pseudomonas spp., and Enterococcus in 1 patient. CONCLUSION: The cultures from samples obtained at discharge showed the presence of antibiotic-resistant pathogenic microorganisms causing healthcare associated infection. KEYWORDS: Child hospitalized, hand hygiene, healthcare associated infection