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The increased  Salmonella  resistance to quinolones and fluoroquinolones is a public health concern in the Southeast Asian region. The objective of this study is to develop a high resolution melt curve (HRM) assay to rapidly screen for mutations in quinolone-resistant determining region (QRDR) of gyrase and topoisomerase IV genes. DNA sequencing was performed on 62  Salmonella  strains to identify mutations in the QRDR of  gyrA ,  gyrB ,  parC , and  parE  genes. Mutations were detected in QRDR of  gyrA  ( n  = 52; S83F, S83Y, S83I, D87G, D87Y, and D87N) and  parE  ( n  = 1; M438I).  Salmonella  strains with mutations within QRDR of  gyrA  are generally more resistant to nalidixic acid (MIC 16 > 256  μ g/mL). Mutations were uncommon within the QRDR of  gyrB ,  parC , and  parE  genes. In the HRM assay, mutants can be distinguished from the wild-type strains based on the transition of melt curves, which is more prominent when the profiles are displayed in difference plot. In conclusion, HRM analysis allows for rapid screening for mutations at the QRDRs of gyrase and topoisomerase IV genes in  Salmonella . This assay markedly reduced the sequencing effort involved in mutational studies of quinolone-resistance genes.

High Resolution Melting Analysis for Rapid Mutation Screening in Gyrase and Topoisomerase IV Genes in Quinolone-ResistantSalmonella enterica