Background/aims: It has been shown that quinolone resistance arises due to mutations in the quinolone resistance-determining regions of the drug targets. This study aimed to optimise a multiplex PCR assay to track plasmid-mediated low-level quinolone resistance profiles. Subjects and methods: A multiplex PCR-based-method in which the primers were already established by our team. About 44 samples were collected from 44 patients who enrolled in this study by using surgical site infection (SSI). Results: By targeting the conserved domains of qnrA, qnrB, qnrS and the qnrVC gene families, the primer number was reduced significantly to only four pairs in one multiplex PCR. Using multiplex PCR, 3/44 SSI samples were found to be carrying fluoroquinoloneresistance genes (qnrA, qnrB, qnrS, qnrVC). Conclusion: A multiplex PCR for detecting pathogens as well as identifying quinolone resistance genes all in one reaction was successfully established.