An Increase in the Clinical Isolation of Acquired AmpC beta-Lactamase-Producing Klebsiella pneumoniae in Korea from 2007 to 2010

We investigated the occurrence and genetic basis of AmpC beta-lactamase (AmpC)-mediated antibiotic resistance, by examining Escherichia coli, Klebsiella pneumoniae, and Proteus mirabilis isolates at a university hospital, from 2007 to 2010. The ampC genes were detected by multiplex AmpC PCR, and AmpC-positive strains were subjected to DNA sequencing. Extended-spectrum beta-lactamase (ESBL) production was assessed using the ESBL disk test based on the utilization of boronic acid. Carbapenem-resistant isolates were further investigated by the modified Hodge test, a carbapenemase inhibition test and SDS-PAGE experiments. AmpC expression was detected in 1.6% of E. coli (39 DHA-1, 45 CMY-2, and 1 CMY-1) isolates, 7.2% of K. pneumoniae (39 DHA-1, 45 CMY-2, and 1 CMY-1) isolates, and 2.5% of P. mirabilis (8 CMY-2 and 1 CMY-1) isolates. Of the 198 acquired AmpC producers, 58 isolates (29.3%) also produced an ESBL enzyme. Among the acquired AmpC-producing K. pneumoniae isolates, the minimum inhibitory concentration (MIC) MIC50/MIC90 values for cefoxitin, cefotaxime, cefepime, imipenem, and meropenem were >32/>32, 16/>32, 1/16, 0.25/0.5, and or =2 microg/mL for 2 K. pneumoniae isolates, both of which carried the blaDHA-1 gene with a loss of OmpK36 expression, but were negative for carbapenemase production. The acquisition of AmpC-mediated resistance in K. pneumoniae isolates increased, as did the proportion of AmpC and ESBL co-producers among the hospital isolates. The accurate identification of isolates producing AmpCs and ESBLs may aid in infection control and will assist physicians in selecting an appropriate antibiotic regimen.

First Outbreak of Multidrug-Resistant Klebsiella pneumoniae Producing both SHV-12-Type Extended-Spectrum beta-Lactamase and DHA-1-Type AmpC beta-Lactamase at a Korean Hospital

PURPOSE: Coexistence of different classes of beta-lactamases in a single bacterial isolate may pose diagnostic and therapeutic challenges. We investigated a spread of Klebsiella pneumoniae isolates co-producing an AmpC beta-lactamase and an extended-spectrum beta-lactamase (ESBL) in a university hospital. MATERIALS AND METHODS: Over a three-month period, a total of 11 K. pneumoniae isolates, which exhibited resistance to cefotaxime, aztreonam, and cefoxitin, were isolated. These isolates showed positive to ESBLs by double disk tests. Minimal inhibitory concentrations (MICs) were determined by broth microdilution testing. All isolates were examined by isoelectric focusing, PCR and sequence analysis to identify bla(SHV) and bla(DHA), and molecular typing by pulsed-field gel electrophoresis (PFGE). RESULTS: All 11 isolates were highly resistant (MIC, > or = 128microngram/ml) to ceftazidime, aztreonam, and cefoxitin, while they were susceptible (MIC, < or = 2microngram/ml) to imipenem. The bla(SHV-12) and bla(DHA-1) genes were detected by PCR and sequence analysis. PFGE revealed a similar pattern in 10 of the 11 strains tested. CONCLUSION: This is the first outbreak report of K. pneumoniae in Korea which co-produced SHV-12 and DHA-1 beta-lactamase, and we suggest a clonal spread of multidrug-resistant K. pneumoniae at a hospital.

Genotype distribution of extended-spectrum and AmpC ?-lactamases produced by Escherichia coli and Klebsiella pneumoniae in 10 teaching hospitals of China / 中国感染与化疗杂志

Objective To investigate the genotype distribution of extended-spectrum?-lactamases(ESBLs) and AmpC?-lacta- mases produced by Escherichia coli and Klebsiella pneumoniae in 10 teaching hospitals of China.Methods 90 clinical strains of E.coli and 61 strains of K.pneumoniae isolated in 2003 and confirmed to produce ESBLs were collected from 10 teaching hos- pitals in China.Analytical isoelectric focusing was used to measure the pI of the?-lactamases.Conjugation experiment was used to study the transfer of cefoxitin resistance.Plasmid-mediated AmpC enzyme genes were amplified and sequenced by multiplex polymerase chain reaction (MPCR).Results The prevalence of ESBL-producing E.coli and K.pneumoniae was about 50% in Wuhan,Nanjing and Jinan.The prevalence of ESBL-producing E.coli was lower than K.pneumoniae in Beijing.However,in other hospitals the prevalence of ESBL-producing E.coli was a little higher than K.pneumoniae.About 24.4% of ESBL-pro- ducing E.coli isolates and 19.4% of ESBL-producing K.pneumoniae isolates were resistant to cefoxitin.Cefoxitin-resistant i solate was identified in all hospitals except Shenyang.Major genotype of ESBL-producing isolates was CTX-M.The CTX-M-9 group was the most common group,followed by CTX-M-1.More K.pneumoniae isolates produced both ESBLs and AmpC en- zyme than E.coli.The genotype was CTX-M/DHA-1.The PCR results of 3 transconjugants producing both ESBLs and AmpC enzyme were the same as their donor isolates.Conclusions The genotype of ESBL-producing isolates is mainly CTX-M-9 group in these teaching hospitals.More K.pneumoniae isolates produced both ESBLs and AmpC enzyme than E.coli.Most of these isolates are due to geno type CTX-M/DHA-1,which can spread through plasmid.

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 and Wide Dissemination of CTX-M-type ESBLs, and CMY-2- and DHA-1-type AmpC beta-Lactamases in Korean Respiratory Isolates of Klebsiella pneumoniae

Respiratory isolates of Klebsiella pneumoniae in Korea during 2002-2003 were studied to determine the prevalence and types of extended-spectrum beta-lactamases (ESBLs) and plasmid-mediated AmpC beta-lactamases (PABLs). ESBL-production was tested by double-disk synergy, and genotypes of beta-lactamases were determined by PCR and sequencing. ESBLs were detected in 28.4% of 373 isolates, and the most prevalent types were SHV-12 (63 isolates) and CTX-M-14 (9 isolates). Forty of 75 ESBL-producers (53.5%) also had PABLs: 21 isolates with CMY-2-like, 17 with DHA-1-like. Pulsed-field gel electrophoresis showed 19 types and 25 of 74 isolates had an identical pattern, indicating nosocomial spread. Dissemination of ESBL- and PABL-producing K. pneumoniae strains in Korea is a particular concern, as it limits the choice of antimicrobial agents for treatment of infections.

Isolation of Klebsiella Oxytoca Clinical Isolate Producing AmpC beta-lactamase, DHA-1 / 대한임상미생물학회지

BACKGROUND: Klebsiella oxytoca strain exhibiting an unusual inducible beta-lactam resistance phenotype was isolated from a wound specimen of a patient at a university hospital in August 2002. The isolate was resistant to ampicillin, ampicillin-sulbactam, cephalothin, cefoxitin, and demonstrated reduced inhibition zone diameters for ceftazidime in combination with clavulanate versus those for ceftazidime when tested alone. METHODS: Antimicrobial susceptibilities were tested using the Etest and disk diffusion method. AmpC beta-lactamase production was determined by modified Hodge test. The disk antagonism method was used to detect inducibility of beta-lactamase. Conjugation experiments were performed by the filter mating method using the recipient Escherichia coli J53 Azir strain. PCR and DNA sequencing of DHA-specific PCR products were tested. RESULTS: The double disk synergy test was negative and the modified Hodge test was positive for the K. oxytoca isolate. Antagonism was observed between cefoxitin and oxyimino-cephalosporins. Sequence analysis of the DHA-specific PCR products revealed that they were identical to the amino acid sequence of the DHA-1 beta-lactamase. Transfer of the resistance by conjugation experiments was successful. CONCLUSIONS: We found a plasmid-mediated DHA-1 beta-lactamase-producing K. oxytoca possessing an unusual inducible beta-lactam resistance phenotype was found in a university hospital in Korea. The resistance phenotype was conferred by DHA-1 encoded by a self-transferable plasmid.