A Korean Nationwide Surveillance Study for Non-Typhoidal Salmonella Isolated in Humans and Food Animals from 2006 to 2008: Extended-Spectrum beta-Lactamase, Plasmid-Mediated AmpC beta-Lactamase, and Plasmid-Mediated Quinolone Resistance qnr Genes / 대한임상미생물학회지

BACKGROUND: The emergence of non-typhoidal Salmonella (NTS) with decreased susceptibilities to fluoroquinolone, ampicillin, or ceftriaxone has been reported worldwide. However, current surveillance studies of resistance among NTS in Korea are limited. Thus, the antimicrobial susceptibilities; resistance mechanisms such as extended-spectrum beta-lactamase (ESBL), plasmid-mediated AmpC beta-lactamase (PABL), and plasmid-mediated quinolone resistance (PMQR); and molecular epidemiologic characteristics were investigated in the present study. METHODS: National Institute of Health and National Veterinary Research and Quarantine Service collected NTS strains from 219 clinical and 293 non-clinical specimens from 2006 to 2008. The antimicrobial susceptibilities were determined using the Clinical and Laboratory Standards Institute disk diffusion test. ESBL, PABL, and qnr genotyping were performed using PCR and nucleotide sequencing. Pulsed-field gel electrophoresis was used for the molecular epidemiologic study. RESULTS: The resistance to ampicillin in clinical and non-clinical NTS was 49% and 18 to 47%, respectively. The resistance rates to trimethoprim-sulfamethoxazole in clinical and non-clinical NTS were 8% and 0 to 41%, respectively. The rates to extended-spectrum cephalosporin were 0 to 1%. One CTX-M-15-producing isolate and four CMY-2-producing isolates were detected. Notably, PFGE analysis showed four isolates carrying bla CMY-2, including one non-clinical strain had high clonality. Although the rate of ciprofloxacin resistance was very low, two qnrS1-carrying NTS strains were detected in non-clinical specimens. CONCLUSION: The resistance rates to ampicillin in both clinical and non-clinical NTS were high, while those to trimethoprim-sulfamethoxazole varied depending on the specimen. NTS strains harboring CTX-M-15-type ESBL or CMY-2-type PABL were detected even though the resistance rates to cephalosporins were very low. Four NTS strains carrying the blaCMY-2-gene implied zoonotic infection. Continuous effort to minimize transfer of resistance genes in NTS is necessary.

Use of Boronic Acid Disks for the Detection of Extended-spectrum beta-lactamase and AmpC beta-lactamase in Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca and Proteus mirabilis / 대한임상미생물학회지

BACKGROUND: Accurate detection of organisms producing extended-spectrum beta-lactamase (ESBL) and AmpC beta-lactamase is very important for treatment of patients. However, unlike the ESBL confirmatory test, there are no guidelines for detection of organisms producing AmpC beta-lactamase. We evaluated a detection method using boronic acid (BA) for ESBL and AmpC beta-lactamase. METHODS: Clinical isolates of Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, and Proteus mirabilis showing intermediate resistance or resistance to cefoxitin (FOX) or positive for ESBL were tested. A > or =5 mm increase in zone diameter of ceftazidime/clavulanic acid/BA (CAZ/CA/BA) and/or cefotaxime/clavulanic acid/BA (CTX/CA/BA) versus CAZ/BA and/or CTX /BA was considered positive for ESBL. Likewise, a > or =5 mm increase in zone diameter of FOX/BA and/or cefotetan/BA (CTT/BA) versus FOX and/or CTT alone was considered positive for AmpC beta-lactamase. RESULTS: Among 622 clinical isolates, ESBL positive rates by the CLSI ESBL confirmatory test or by the BA method were 18.1% or 18.4% for E. coli, 38.3% or 40.4% for K. pneumoniae, 8.7% or 8.7% for K. oxytoca, and 14.8% or 14.8% for P. mirabilis, respectively. AmpC beta-lactamase positive rates using the BA method were 3.7% for E. coli, 33.3% for K. pneumoniae, 0% for K. oxytoca, and 7.4% for P. mirabilis. The detection rates of coproducing ESBL and AmpC beta-lactamase were 2.4% in E. coli 27.1% in K. pneumoniae, and 3.7% in P. mirabilis. CONCLUSION: The ESBL confirmatory method using BA was found to enhance the detection of ESBLs, even when potentially masked by AmpC beta-lactamase.

Epidemiology of Plasmid-Mediated AmpC beta-Lactamase Produced by Clinical Isolates of Klebsiella pneumoniae from Four University Hospitals in Korea / 병원감염관리

BACKGROUND: Plasmid-mediated AmpC beta-lactamases (PABLs) are cephalosporinases that confer resistance to a wide variety of beta-lactam drugs and that may thereby create serious therapeutic problems. The PABL-producing organisms are a major concern in nosocomial infections and should therefore be monitored in surveillance studies. METHODS: During the period of May to July 2004, 27 cefoxitin non-susceptible isolates of Klebsiella pneumoniae from four university hospitals (Seoul 2, Daejeon 1, and Choongju 1) were tested for antimicrobial susceptibility by the broth microdilution method. The cefoxitin non-susceptible isolates were further investigated by the double disk synergy test for extended-spectrum beta-lactamases, multiplex AmpC PCR, DNA sequencing, and pulsed-field gel electrophoresis (PFGE). RESULTS: PABL-producing K. pneumoniae were found in all the four hospitals. Eight (32%) of 25 PABL producers were also tested positive by double disk synergy tests. Susceptibilities of the PABL producers were as follows: ceftazidime, 4%; aztreonam, 36%; cefepime, 76%; and imipenem, 100%. Among the 25 K. pneumoniae isolates were 24 DHA-1 and 1 CMY-1 beta-lactamase producers. The PFGE patterns of the DHA-1-producing K. pneumoniae showed variable as well as identical patterns. CONCLUSION: PABL-producing K. pneumoniae is widespread among medical institutions in Korea. A DHA-1 type in K. pneumoniae was the predominant enzyme detected. Overall, despite many different PFGE patterns of the PABL producers, some outbreak and epidemic clones appear to be prevalent in some hospitals in Korea. For the prevention of the spread of PABL-producing K. pneumoniae, it should be identified accurately by the clinical laboratory.

Emergence of Plasmid-Mediated CMY-2 beta -Lactamase Produced by Clinical Isolates of Escherichia coli in Korea / 대한진단검사의학회지

BACKGROUND: Of the plasmid-mediated AmpC beta-lactamases (ABLs), CMY-2 is the most prevalent and is distributed in many countries. However, little is known about the emergence and characteristics of CMY-2 among Escherichia coli isolates in Korea. The aims of this study were to detect the emergence of the CMY-2 beta-lactamase in clinical isolates of E. coli from various regions in Korea. METHODS: Eighteen cefoxitin non-susceptible isolates of 1, 130 consecutive, nonrepeat isolates of E. coli at five university hospitals were tested for antimicrobial susceptibility by the broth microdilution method. The cefoxitin non-susceptible isolates were further investigated by AmpC disk tests, double disk synergy (DDS) tests, isoelectric focusing, CMY-2-specific PCR, DNA sequencing, and plasmid analysis. RESULTS: Seven (0.6%) isolates of plasmid-mediated ABL-producing E. coli were found at three of the five hospitals; all seven isolates produced CMY-2 beta-lactamase and one of the isolates was also tested positive by the DDS test. All isolates demonstrated different plasmid patterns by plasmid analysis. CONCLUSIONS: Our data indicate that CMY-2-producing E. coli has emerged and is prevalent in the medical institution in Korea. Therefore, constant surveillance is needed to prevent its further spread.