The occurrence of resistance to last-resort antibiotics was evaluated among <i>Enterobacteriaceae</i> isolates recovered from hospitalized children in a remote African archipelago, São Tomé and Príncipe, where there is limited access to those antibiotics. Fifty patients were screened for colonization by carbapenem-, pan-aminoglycoside-, or polymyxin-resistant <i>Enterobacteriaceae</i> A total of 36 isolates (including 30 <i>Escherichia coli</i> and 4 <i>Klebsiella pneumoniae</i> ) were recovered from 23 patients, including 26 isolates harboring the <i>bla</i> _OXA-181 carbapenemase gene, a single isolate harboring the 16S rRNA methylase gene <i>rmtB</i> encoding pan-resistance to aminoglycosides, and 8 isolates coharboring both genes. A single isolate possessed the plasmid-borne colistin resistance gene <i>mcr-1</i> A high clonal relationship was found for OXA-181-producing <i>E. coli</i> (4 clones), and conversely, three of the four OXA-181-producing <i>K. pneumoniae</i> isolates were clonally unrelated. This study overall showed a high prevalence of resistance to last-resort antibiotics in this country, where no epidemiological data were previously available.