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Ninety-three Malaysian extended-spectrum  β -lactamase (ESBL)-producing  Klebsiella pneumoniae  isolates were investigated for ciprofloxacin resistance. Two mismatch amplification mutation (MAMA) assays were developed and used to facilitate rapid detection of  gyrA  and  parC  mutations. The isolates were also screened for plasmid-mediated quinolone resistance (PMQR) genes including  aac(6 ′ )-Ib-cr, qepA , and  qnr . Ciprofloxacin resistance (MICs 4– ≥ 32   μ  g/mL) was noted in 34 (37%) isolates, of which 33 isolates had multiple mutations either in  gyrA  alone ( n  = 1) or in both  gyrA  and  parC  regions ( n  = 32).  aac(6 ′ )-Ib-cr  was the most common PMQR gene detected in this study ( n  = 61), followed by  qnrB  and  qnrS  ( n  = 55 and 1, resp.). Low-level ciprofloxacin resistance (MICs 1-2   μ  g/mL) was noted in 40 (43%) isolates carrying  qnrB  accompanied by either  aac(6 ′ )-Ib-cr  ( n  = 34) or a single  gyrA  83 mutation ( n  = 6). Ciprofloxacin resistance was significantly associated with the presence of multiple mutations in  gyrA  and  parC  regions. While the isolates harbouring  gyrA  and/or  parC  alteration were distributed into 11 PFGE clusters, no specific clusters were associated with isolates carrying PMQR genes. The high prevalence of ciprofloxacin resistance amongst the Malaysian ESBL-producing  K. pneumoniae  isolates suggests the need for more effective infection control measures to limit the spread of these resistant organisms in the hospital.

biomed research international

Molecular Analysis of Ciprofloxacin Resistance Mechanisms in Malaysian ESBL-ProducingKlebsiella pneumoniaeIsolates and Development of Mismatch Amplification Mutation Assays (MAMA) for Rapid Detection ofgyrAandparCMutations