© Copyright 2020, Mary Ann Liebert, Inc., publishers 2020.Escherichia coli is the leading etiological agent of community-acquired urinary tract infection (UTI). Fluoroquinolones have long been the choice of empirical treatment for UTIs. Plasmid-mediated fluoroquinolone-resistance (PMFR) is important not only for conferring resistance to fluoroquinolones but also because of the presence of PMFR genes on plasmids carrying genes encoding resistance to other antimicrobials. In this study, we aimed at investigating the frequency of PMFR genes in fluoroquinolone-resistant and/or expanded spectrum beta-lactamase (ESBL)-producing E. coli strains isolated from pediatric and adult patients diagnosed with UTI. E. coli strains isolated from urine cultures of 141 adult and 117 pediatric outpatients were evaluated. Antimicrobial susceptibilities were interpreted according to the EUCAST criteria. The presence of PMFR genes (qnrA, qnrB, qnrC, qnrS, qepA, aac(6′)-Ib, and aac(6′)-Ib-cr) was investigated by multiplex PCR analysis. One hundred-three (73.05%) adult and 92 (78.63%) pediatric isolates were fluoroquinolone resistant and/or ESBL producers. One third (92/258) of all isolates carried at least one PMFR gene, the most prevalent one being qnrS (67.4%). None of the isolates carried qnrC and qepA genes. PMFR determinants were found to be widespread among adult and pediatric isolates. Rational antimicrobial use is crucial for prevention of resistance in both adult and pediatric populations.