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Two mismatch amplification mutation (MAMA) assays were developed and used to facilitate rapid detection of<jats:italic>gyrA<\/jats:italic>and<jats:italic>parC<\/jats:italic>mutations. The isolates were also screened for plasmid-mediated quinolone resistance (PMQR) genes including<jats:italic>aac(6<\/jats:italic>\u2032<jats:italic>)-Ib-cr, qepA<\/jats:italic>, and<jats:italic>qnr<\/jats:italic>. Ciprofloxacin resistance (MICs<mml:math xmlns:mml=\"http:\/\/www.w3.org\/1998\/Math\/MathML\" id=\"M2\"><mml:mn>4<\/mml:mn><mml:mtext>\u2013<\/mml:mtext><mml:mo>\u2265<\/mml:mo><mml:mn>32<\/mml:mn><\/mml:math>\u2009<jats:italic>\u03bc<\/jats:italic>g\/mL) was noted in 34 (37%) isolates, of which 33 isolates had multiple mutations either in<jats:italic>gyrA<\/jats:italic>alone<mml:math xmlns:mml=\"http:\/\/www.w3.org\/1998\/Math\/MathML\" id=\"M3\"><mml:mo stretchy=\"false\">(<\/mml:mo><mml:mi>n<\/mml:mi><mml:mo>=<\/mml:mo><mml:mn>1<\/mml:mn><mml:mo stretchy=\"false\">)<\/mml:mo><\/mml:math>or in both<jats:italic>gyrA<\/jats:italic>and<jats:italic>parC<\/jats:italic>regions<mml:math xmlns:mml=\"http:\/\/www.w3.org\/1998\/Math\/MathML\" id=\"M4\"><mml:mo stretchy=\"false\">(<\/mml:mo><mml:mi>n<\/mml:mi><mml:mo>=<\/mml:mo><mml:mn>32<\/mml:mn><mml:mo stretchy=\"false\">)<\/mml:mo><\/mml:math>.<jats:italic>aac(6<\/jats:italic>\u2032<jats:italic>)-Ib-cr<\/jats:italic>was the most common PMQR gene detected in this study<mml:math xmlns:mml=\"http:\/\/www.w3.org\/1998\/Math\/MathML\" id=\"M5\"><mml:mo stretchy=\"false\">(<\/mml:mo><mml:mi>n<\/mml:mi><mml:mo>=<\/mml:mo><mml:mn>61<\/mml:mn><mml:mo stretchy=\"false\">)<\/mml:mo><\/mml:math>, followed by<jats:italic>qnrB<\/jats:italic>and<jats:italic>qnrS<\/jats:italic>(<mml:math xmlns:mml=\"http:\/\/www.w3.org\/1998\/Math\/MathML\" id=\"M6\"><mml:mi>n<\/mml:mi><mml:mo>=<\/mml:mo><mml:mn>55<\/mml:mn><\/mml:math>and 1, resp.). Low-level ciprofloxacin resistance (MICs 1-2\u2009<jats:italic>\u03bc<\/jats:italic>g\/mL) was noted in 40 (43%) isolates carrying<jats:italic>qnrB<\/jats:italic>accompanied by either<jats:italic>aac(6<\/jats:italic>\u2032<jats:italic>)-Ib-cr<\/jats:italic><mml:math xmlns:mml=\"http:\/\/www.w3.org\/1998\/Math\/MathML\" id=\"M7\"><mml:mo stretchy=\"false\">(<\/mml:mo><mml:mi>n<\/mml:mi><mml:mo>=<\/mml:mo><mml:mn>34<\/mml:mn><mml:mo stretchy=\"false\">)<\/mml:mo><\/mml:math>or a single<jats:italic>gyrA<\/jats:italic>83 mutation<mml:math xmlns:mml=\"http:\/\/www.w3.org\/1998\/Math\/MathML\" id=\"M8\"><mml:mo stretchy=\"false\">(<\/mml:mo><mml:mi>n<\/mml:mi><mml:mo>=<\/mml:mo><mml:mn>6<\/mml:mn><mml:mo stretchy=\"false\">)<\/mml:mo><\/mml:math>. Ciprofloxacin resistance was significantly associated with the presence of multiple mutations in<jats:italic>gyrA<\/jats:italic>and<jats:italic>parC<\/jats:italic>regions. While the isolates harbouring<jats:italic>gyrA<\/jats:italic>and\/or<jats:italic>parC<\/jats:italic>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<jats:italic>K. pneumoniae<\/jats:italic>isolates suggests the need for more effective infection control measures to limit the spread of these resistant organisms in the hospital.<\/jats:p>","DOI":"10.1155\/2014\/601630","type":"journal-article","created":{"date-parts":[[2014,4,10]],"date-time":"2014-04-10T21:03:22Z","timestamp":1397163802000},"page":"1-10","source":"Crossref","is-referenced-by-count":30,"title":["Molecular Analysis of Ciprofloxacin Resistance Mechanisms in Malaysian ESBL-Producing<i>Klebsiella pneumoniae<\/i>Isolates and Development of Mismatch Amplification Mutation Assays (MAMA) for Rapid Detection of<i>gyrA<\/i>and<i>parC<\/i>Mutations"],"prefix":"10.1155","volume":"2014","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5711-6264","authenticated-orcid":true,"given":"Farah","family":"Al-Marzooq","sequence":"first","affiliation":[{"name":"Department of Medical Microbiology, Faculty 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