The spread of plasmid-mediated quinolone resistance determinants QnrA and QnrS was evaluated in a collection of 186 extended-spectrum beta-lactamase (ESBL)-positive enterobacterial isolates from 2002 to 2005 and 185 nalidixic acid-resistant strains isolated during the first 6 months of 2005 at the Bicêtre hospital, France. Out of these 186 ESBL-positive isolates, 2.2 and 1.6% carried a QnrA1 and a QnrS1 determinant, respectively. The ESBLs associated with QnrA1 were VEB-1, SHV-12, and CTX-M-1, whereas those associated with QnrS1 were TEM-52, SHV-12, and CTX-M-1. Among the 185 nalidixic acid-resistant strains isolated in 2005, 0.5 and 2.7% had a QnrA1 determinant and a QnrS1 determinant, respectively. The genetic environments of the qnrA1 gene differed but were always associated with sul1 type integrons. In contrast, qnrS1 genes were not embedded in class 1 integrons but located often (but not systematically) downstream of the insertion sequence ISEcl2 on plasmids that often carried a novel beta-lactamase gene, bla(LAP-1). This is the first study identifying the QnrS resistance determinant in Europe and indicating that this determinant might also be widespread.