blaSHV-12 gene detection from Klebsiella pneumoniae producing Extended-Spectrum β-Lactamase using amplification-refractory mutation system method
Extended-Spectrum β-lactamase (ESBL)-producing bacteria makes the bacterial disease more difficult to treat resulting in increased cost, longer duration of patient treatment along with increased morbidity and mortality. Klebsiella pneumoniae is one such bacteria known to produce ESBL. The objective of this study was to determine the presence of the blaSHV-12 gene in ESBL-producing K. pneumoniae from pneumonia patients at Dr. Hasan Sadikin Teaching Hospital, Bandung, and to determine the relationship of the blaSHV-12 gene with the incidence of antibiotic resistance. The blaSHV-12 gene was detected using Amplification-Refractory Mutation System (ARMS) method and visualized by UV spectroscopy following SYBR® safe DNA gel stain embedded agarose gel electrophoresis. The presence of the blaSHV-12 gene was indicated by the appearance of three bands at 756, 397, and 142 bp. To our knowledge, this is the first study to use the ARMS method with specific tetra primers in search of the blaSHV-12 gene. We collected antibiotic susceptibility data retrospectively from the Department of Clinical Pathology, testing with VITEK 2. Correlation test using phi coefficient tests. From 45 samples of ESBL-producing K. pneumoniae tested in this study, a total of 39 (87%) samples were blaSHV-12 gene-positive while 6 samples (13%) were blaSHV-12 gene negative. Our results indicate that the presence of the blast-12 gene correlates with cefmethazole (value -0,544) but does not correlate with sensitivity to ampicillin/sulbactam, piperacillin/tazobactam, cefoxitin, ertapenem, and meropenem.
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