The trends and types of carbapenemase-producing Gram-negative bacilli were analyzed from clinical specimens collected between 2005 and 2012 at a Korean teaching hospital. The proportions of carbapenem-resistant Acinetobacter spp. increased markedly to 66%. Metallo-beta-lactamase producers significantly decreased and the majority shifted from the bla(VIM-2) type to the bla(IMP-1) type.
Acinetobacter/classification/drug effects/*enzymology ; Acinetobacter Infections/drug therapy ; Anti-Bacterial Agents/*pharmacology ; Bacterial Proteins ; Carbapenems/*pharmacology ; Drug Resistance, Microbial ; Gram-Negative Bacteria/*drug effects/enzymology/isolation & purification ; Humans ; Incidence ; Microbial Sensitivity Tests/trends ; Population Surveillance ; Pseudomonas/classification/drug effects/enzymology ; Republic of Korea/epidemiology ; beta-Lactamases/biosynthesis/*drug effects

BACKGROUND/AIMS: We examined the prevalence of extended-spectrum beta-lactamase (ESBL) production and the impact of ESBL on clinical outcomes in cancer patients with Enterobacter spp. bacteremia. METHODS: Using prospective cohort data on Enterobacter bacteremia obtained between January 2005 and November 2008 from a tertiary care center, the prevalence and clinical impact of ESBL production were evaluated. RESULTS: Two-hundred and three episodes of Enterobacter spp. bacteremia were identified. Thirty-one blood isolates (15.3%, 31/203) scored positive by the double-disk synergy test. Among 17 isolates in which ESBL genes were detected by polymerase chain reaction and sequencing, CTX-M (n = 12), SHV-12 (n = 11), and TEM (n = 4) were the most prevalent ESBL types. Prior usage of antimicrobial agents (77.4% vs. 54.0%, p = 0.02) and inappropriate empirical antimicrobial therapy (22.6% vs. 3.0%, p < 0.001) were more commonly encountered in the ESBL-positive group than in the extended-spectrum cephalosporin-susceptible ESBL-negative group, respectively. Clinical outcomes did not differ significantly between the two groups (30-day mortality rate, 19.4% vs. 17.0%, p = 0.76; median length of hospital stay, 24.0 days vs. 30.5 days, p = 0.97). Initial presentation of severe sepsis/septic shock, pneumonia, and intra-abdominal infection were independently associated with 30-day mortality. CONCLUSIONS: The prevalence of ESBL-producing isolates was 15.3% in cancer patients with Enterobacter bacteremia. Although inappropriate empirical therapy was more common in the ESBL-positive group, ESBL production was not associated with poorer outcomes.
Adult ; Aged ; Anti-Bacterial Agents/therapeutic use ; Bacteremia/*complications/drug therapy/microbiology ; Child ; Cohort Studies ; Enterobacter/*enzymology/genetics ; Enterobacteriaceae Infections/*complications/drug therapy/microbiology ; Female ; Humans ; Infant ; Male ; Middle Aged ; Neoplasms/*complications ; Prospective Studies ; Treatment Outcome ; beta-Lactamases/*biosynthesis/genetics

OBJECTIVETo identify risk factors for Klebsiella pneumoniae carbapenemase-producing Klebsiella pneumoniae (KPC-Kp) colonization in neonates hospitalized in the neonatal intensive care unit (NICU).

METHODSA case-control study was conducted. The case group included nine patients colonized with KPC-Kp between 1 August 2012 and 31 April 2013 and the controls were selected randomly from patients without KPC-Kp colonization during the same period. Univariable analysis and multivariable logistic regression analysis were conducted to identify risk factors for KPC-Kp colonization.

RESULTSThe univariable analysis showed 11 factors associated with KPC-Kp colonization: gestational age, birth weight, length of hospital stay, duration of mechanical ventilation, congenital heart disease, peripherally inserted central catheter, surgical operation, duration of intravenous nutrition, carbapenems use, duration of carbapenems use and glycopeptides use. The multivariable logistic regression analysis showed that exposure to more than 4 days of carbapenems use (OR=18.7, 95%CI: 1.98-175.5, P=0.01) was an independent risk factor for KPC-Kp colonization. The intervention to control KPC-Kp colonization included contact isolation, active surveillance, and rational use of antibiotics.

CONCLUSIONSExposure to prolonged use of carbapenems is an independent risk factor for the development of KPC-Kp colonization in neonates hospitalized in the NICU.

Bacterial Proteins ; biosynthesis ; Carbapenems ; adverse effects ; Female ; Humans ; Infant, Newborn ; Intensive Care Units, Neonatal ; Klebsiella pneumoniae ; enzymology ; isolation & purification ; Logistic Models ; Male ; Risk Factors ; beta-Lactamases ; biosynthesis

OBJECTIVETo evaluate the antimicrobial activity of total alkaloids extracted from Sophorea alopecuroides L. (TASA) against clinical isolated extended-spectrum beta-lactamases (ESBLs) producing Escherichia coli (E. coli) strains.

METHODSThe antibacterial activity of TASA either itself or in combination with cefotaxime (CTX) or ceftazidime (CAZ) was investigated by using the microbroth dilution method and phenotypic confirmatory disk diffusion test against three clinical isolated ESBLs-producing E. coli strains; the interactions of TASA and CTX or CAZ were ascertained by evaluating the fractional inhibitory concentration index (FICI).

RESULTSThe antibacterial activity of either TASA itself or in combination with CTX or CAZ was found. The minimum inhibitory concentration (MICs) of TASA against the ESBLs producing isolates was 12.5 mg/mL. In the combinations with a sub-inhibitory concentration of TASA, a synergistic effect on CTX and CAZ against the ESBLs producing isolates was observed. Similarly, the isolates exposed to lower dose of TASA yielded an increased susceptibility to CTX and CAZ by 8-16 folds determined by microdilution assay. Moreover, enzymatic detection of ESBLs demonstrated that TASA induced reversal resistance to CTX and CAZ partially by a mechanism of inhibition of ESBLs activity in these isolates. Additionally, in the tested isolates following the exposure of TASA, molecular analysis verified the SHV-type beta-lactamase encoding ESBL gene in these isolates, and no mutation was introduced into the ESBL gene.

CONCLUSIONSThese results suggest that TASA could be used as a source of natural compound with pharmacological activity of reversal resistance to antimicrobial agent. These findings also indicated that the application of the TASA in combination with antibiotics might prove useful in the control and treatment of infectious diseases caused by the ESBLs producing enterobacteriaceae.

Alkaloids ; isolation & purification ; pharmacology ; Anti-Bacterial Agents ; pharmacology ; Base Sequence ; Cefotaxime ; pharmacology ; Ceftazidime ; pharmacology ; DNA Primers ; Escherichia coli ; drug effects ; enzymology ; Microbial Sensitivity Tests ; Polymerase Chain Reaction ; Sophora ; chemistry ; beta-Lactamases ; biosynthesis

OBJECTIVETo investigate the characteristics of New Delhi metallo-beta-lactamase-1 (NDM-1) gene of Klebsiella pneumoniae (K. pneumoniae) emerging in Hunan, and its relationship to antibiotic resistance.

METHODSThe clinical strain was isolated from a sputum sample of a child with severe pnemonia and toxic myocarditis who was admitted into a general hospital of Hunan Province. VITEK-2 compact instrument was used for bacterial identification and antibiotic susceptibility test. Modified Hodge test was used for the screening of carbapenemase. EDTA-synergy test and combination disk diffusion test were used for detection of metallo-&beta;-lactamase (MBL). PCR was performed for amplification of NDM-1 genes and the positive products were sequenced and analyzed with BLAST. Conjugation was also performed to analyze mechanisms of antibiotic resistance. The results of antibiotic susceptibility tests were compared before and after conjugation.

RESULTSThe isolated strain was identified as K.pneumoniae. Modified Hodge test, EDTA-synergy test and combination disk diffusion test were all positive for the strain. The homology between gene sequence of PCR amplification products and NDM-1 gene FN396876.1 in the GenBank was 100%. Transconjugant DNA was used as template for the amplification of NDM-1 gene. The amplification products were sequenced and found to be the same as the NDM-1 gene amplification product of the donor strain. The MIC of transconjugant E.coli J53 (NDM-1) to all the &beta;-lactams increased significantly compared with the recipient strain E.coli J53. The MIC of ertapenem and imipenem increased by more than 8 times, while the MIC of ceftazidime and ceftriaxone increased by more than 64 times.

CONCLUSIONSThis study first identified a strain of K. pneumoniae carrying NDM-1 in mainland China. NDM-1 gene can be transmitted among different strains and causes extensively drug-resistance to &beta;-lactams.

Base Sequence ; China ; Drug Resistance, Bacterial ; Klebsiella pneumoniae ; drug effects ; enzymology ; genetics ; Molecular Sequence Data ; Polymerase Chain Reaction ; beta-Lactamases ; biosynthesis ; genetics

OBJECTIVETo study the prevalence and drug resistance of extended-spectrum-&beta;-lactamases (ESBLs)-producing bacteria in blood culture isolated from children with hematological malignancy after chemotherapy.

METHODSBlood samples taken from 3264 children with hematological malignancy and severe infection following chemotherapy between 2002 and 2008 were cultured using the Bact/ALTER 3D blood culture system. VITEK 60 automicroscan was used to identify viral species and to conduct drug resistance tests. The results were indentified according to National Committee for Clinical Laboratory Standard guidelines.

RESULTSFifty-eight strains of Escherichia coli and fifty-one strains of Klebsiella pneumoniae were isolated. Thirty-eight strains of Escherichia coli and nineteen strains of Klebsiella pneumoniae were ESBLs-producing and these ESBLs-producing strains were less susceptible than those that were non-ESBLs-producing to most antibiotics. Both ESBL- and non-ESBL-producing strains were susceptible to imipenem, piperacillin/tazobactam and amikacin.

CONCLUSIONSThe prevalence of ESBLs-producing bacteria is high in childrn with hematological malignancy and infection following chemotherapy. ESBLs-producing bacteria are resistant to common antibiotics, suggesting that antibiotic treatment based on the result of antimicrobial susceptibility test is necessary in these children.

Adolescent ; Bacteremia ; microbiology ; Child ; Child, Preschool ; Drug Resistance, Bacterial ; Escherichia coli ; drug effects ; enzymology ; isolation & purification ; Female ; Hematologic Neoplasms ; drug therapy ; microbiology ; Humans ; Klebsiella pneumoniae ; drug effects ; enzymology ; isolation & purification ; Male ; Microbial Sensitivity Tests ; beta-Lactamases ; biosynthesis

BACKGROUND/AIMS: We evaluated the clinical features of ciprofloxacin-resistant Proteus mirabilis bacteremia and risk factors for ciprofloxacin resistance. METHODS: From October 2000 to July 2009, 37 patients with clinically significant P. mirabilis bacteremia were identified and data from patients with ciprofloxacin-resistant and ciprofloxacin-susceptible P. mirabilis bacteremia were compared. RESULTS: The most common underlying diseases were neurologic disease (37.8%) and solid tumors (29.7%). The most common site of infection was the urinary tract (35.1%). Ten of the 37 patients (27.0%) were infected with ciprofloxacin-resistant isolates, and univariate analysis revealed a significant relationship between ciprofloxacin-resistant P. mirabilis bacteremia and neurologic disease, recent operation, L-tube insertion, percutaneous tube use, and extended-spectrum beta-lactamase (ESBL) production (all p < 0.05). ESBL was detected in six of 10 (60%) ciprofloxacin-resistant isolates, while only three of 27 (11%) ciprofloxacin-susceptible isolates produced ESBL (p = 0.005). In a logistic regression analysis, ESBL production remained a significant factor associated with ciprofloxacin resistance, after adjusting for other variables. CONCLUSIONS: These data indicate a close association between ciprofloxacin resistance and ESBL-production in P. mirabilis bacteremia. This association is particularly troublesome because the therapeutic options for serious infections caused by ESBL-producing P. mirabilis are severely restricted.
Adult ; Aged ; Anti-Infective Agents/*pharmacology ; Bacteremia/*drug therapy ; Ciprofloxacin/*pharmacology ; Drug Resistance, Bacterial ; Female ; Humans ; Male ; Middle Aged ; Proteus Infections/*drug therapy ; Proteus mirabilis/*drug effects/enzymology ; Risk Factors ; beta-Lactamases/*biosynthesis

OBJECTIVETo study the risk factors for infection with extended-spectrum beta-lactamase (ESBL) producing strains in children.

METHODSThe clinical data of 242 pediatric in-patients with lower respiratory tract infections from February 2009 to January 2011 were retrospectively analyzed. The risk factors of ESBL-producing strain infections were investigated using univariate analysis and multivariate logistic regression analysis.

RESULTSUnivariate analysis showed that six factors were related with ESBL-producing strain infections: repeated sucking of phlegm (OR: 2.279, P<0.01), tracheal intubation(OR: 3.101, P<0.01), administration of the third generation cephalosporin for more than three days (OR: 3.628, P<0.01), admission to the pediatric intensive care unit (PICU) (OR: 2.378, P<0.01), indwelling of nasogastric tube (OR: 2.460, P<0.01), prophylactic use of antibiotics (OR: 1.747, P<0.05). Multivariate logistic regression showed that the application of the third-generation cephalosporin for more than three days (OR: 5.672, P<0.01), repeated sucking of phlegm (OR: 3.917, P<0.01), tracheal intubation (OR: 3.717, P<0.01), indwelling of nasogastric tube (OR: 2.961, P<0.01), and admission to PICU (OR: 3.237, P<0.01) were the independent risk factors for the infections.

CONCLUSIONSThe infections of ESBL-producing strains are caused by many factors, among which the application of the third-generation cephalosporin for more than three days, invasive operations, and admission to PICU are the independent risk factors.

Adolescent ; Bacterial Infections ; etiology ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Risk Factors ; beta-Lactamases ; biosynthesis

Carbapenem-resistant Klebsiella pneumoniae isolates producing K. pneumoniae carbapenemases (KPC) were first reported in the USA in 2001, and since then, this infection has been reported in Europe, Israel, South America, and China. In Korea, the first KPC-2-producing K. pneumoniae sequence type (ST) 11 strain was detected in 2010. We report the case of a patient with a urinary tract infection caused by KPC-2-producing K. pneumoniae. This is the second report of a KPC-2-producing K. pneumoniae infection in Korea, but the multilocus sequence type was ST258. The KPC-2-producing isolate was resistant to all tested beta-lactams (including imipenem and meropenem), amikacin, tobramycin, ciprofloxacin, levofloxacin, and trimethoprim-sulfamethoxazole, but was susceptible to gentamicin, colistin, polymyxin B, and tigecycline. The KPC-2-producing isolate was negative to phenotypic extended-spectrum beta-lactamase (ESBL) and AmpC detection tests and positive to modified Hodge test and carbapenemase inhibition test with aminophenylboronic acid.
Aged ; Bacterial Proteins/antagonists & inhibitors/metabolism ; Carbapenems/pharmacology ; Drug Resistance, Bacterial/genetics ; Female ; Humans ; Klebsiella pneumoniae/drug effects/genetics/*isolation & purification ; Microbial Sensitivity Tests ; Republic of Korea ; Sequence Analysis, DNA ; Urinary Tract Infections/*diagnosis/microbiology ; beta-Lactamases/antagonists & inhibitors/biosynthesis/*genetics/metabolism

BACKGROUND: Members of the Acinetobacter calcoaceticus-baumannii (Acb) complex are important opportunistic bacterial pathogens and present significant therapeutic challenges in the treatment of nosocomial infections. In the present study, we investigated the integrons and various genes involved in resistance to carbapenems, aminoglycosides, and fluoroquinolones in 56 imipenem-nonsusceptible Acb complex isolates. METHODS: This study included 44 imipenem-nonsusceptible A. baumannii, 10 Acinetobacter genomic species 3, and 2 Acinetobacter genomic species 13TU strains isolated in Daejeon, Korea. The minimum inhibitory concentrations (MICs) were determined by Etest. PCR and DNA sequencing were used to identify the genes that potentially contribute to each resistance phenotype. RESULTS: All A. baumannii isolates harbored the blaOXA-51-like gene, and 21 isolates (47.7%) co-produced OXA-23. However, isolates of Acinetobacter genomic species 3 and 13TU only contained blaIMP-1 or blaVIM-2. Most Acb complex isolates (94.6%) harbored class 1 integrons, armA, and/or aminoglycoside-modifying enzymes (AMEs). Of particular note was the fact that armA and aph(3')-Ia were only detected in A. baumannii isolates, which were highly resistant to amikacin (MIC50> or =256) and gentamicin (MIC50> or =1,024). In all 44 A. baumannii isolates, resistance to fluoroquinolones was conferred by sense mutations in the gyrA and parC. However, sense mutations in parC were not found in Acinetobacter genomic species 3 or 13TU isolates. CONCLUSIONS: Several differences in carbapenem, aminoglycoside, and fluoroquinolone resistance gene content were detected among Acb complex isolates. However, most Acb complex isolates (87.5%) possessed integrons, carbapenemases, AMEs, and mutations in gyrA. The co-occurrence of several resistance determinants may present a significant threat.
Acinetobacter Infections/microbiology ; Acinetobacter baumannii/*genetics/isolation & purification ; Anti-Bacterial Agents/*pharmacology ; Bacterial Proteins/genetics ; DNA Gyrase/genetics ; DNA, Bacterial/chemistry/genetics ; Drug Resistance, Bacterial/*genetics ; Humans ; Imipenem/*pharmacology ; Integrons/genetics ; Methyltransferases/genetics ; Microbial Sensitivity Tests ; Mutation ; Polymerase Chain Reaction ; Republic of Korea ; Sequence Analysis, DNA ; beta-Lactamases/biosynthesis/genetics