Antimicrobial resistance (AMR) is a concern in medical care for children who have high burden of infectious diseases. We investigated the prevalence of bacterial species and their susceptibility to antimicrobials of 1019 clinical isolates from pediatric patients in a tertiary-care hospital in Yangon, Myanmar for one-year period (2020). The most frequently recovered species was Escherichia coli, followed by Klebsiella pneumoniae and Staphylococcus aureus, all of which accounted for 43% of clinical isolates, while 25% of isolates comprised non-fermenter, including Pseudomonas sp. and Acinetobacter sp. Phenotypically determined ESBL (extended-spectrum beta-lactamase)-positive rates in E. coli, K. pneumoniae, and Enterobacter sp. were 82%, 88%, and 65%, respectively. High rates of multiple drug resistance were noted for E. coli (84%), K. pneumoniae (81%), and Acinetobacter sp. (65%), associated with carbapenem resistance in 48%, 42%, and 59% of isolates, respectively. In contrast, S. aureus isolates exhibited low resistance rates (<30%) to most of antimicrobials, with 22% being resistant to oxacillin/cefoxitin. Fluoroquinolone resistance was found in most of bacterial species with different prevalence rates. The present study revealed the current status on prevalence of bacterial species causing infections in pediatric patients in Myanmar, highlighting the significance to monitor AMR among children.