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[Activity of fosfomycin against extended-spectrum beta-lactamase producing Escherichia coli and Klebsiella pneumoniae].

INTRODUCTION: Infection due to extended-spectrum beta-lactamase (ESBL)-producing microorganisms is an emerging problem in the community; a high proportion of these microorganisms have been isolated from urine samples of women with uncomplicated urinary tract infections (UTI). The options for oral treatment of uncomplicated UTI are limited because of the multiple drug resistance typical of ESBL-producing strains. METHODS: The in vitro activity of fosfomycin (FOS) was determined against 428 ESBL-producing strains, including 290 (68%) E. coli and 138 (32%) K. pneumoniae. Activity of fosfomycin was compared with that of amoxicillin-clavulanate (AMC), ciprofloxacin (CIP) and cotrimoxazole (SxT). MICs of AMC, CIP, and SxT, and detection of ESBL production were tested by the broth microdilution method, whereas FOS MICs were determined by the agar dilution method. ESBLs were characterized by isoelectric focusing, polymerase chain reaction (PCR) and direct sequencing of encoding genes. The genetic relationship among the isolates was determined by REP-PCR. RESULTS: Among the 428 ESBL-producing isolates studied, 417 (97.4%) were susceptible to FOS (MIC < or = 64 microg/mL). The resistance rate of E. coli to FOS was 0.3%, and was lower than resistance to AMC (11.7%), whereas the resistance rate of K. pneumoniae was 7.2% and was equal to resistance to AMC. SxT and CIP were the least active antibiotic agents against ESBL-producing isolates (sensitivity < 50%). There were no differences in fosfomycin activity against strains expressing different types of ESBLs. CONCLUSION: Fosfomycin showed maintained activity against ESBL-producing strains and did not present co-resistance with other antimicrobial groups.

Amoxicillin-Potassium Clavulanate Combination↗

Antimicrobial resistance surveillance of gram-negative bacteria isolated from intensive care units of four different hospitals in Turkey. Evaluation of the prevalence of extended-spectrum and inducible beta-lactamases using different E-test strips and direct induction methods.

BACKGROUND: The aim of this study was to determine the antimicrobial resistance of gram-negative rods (GNRs) isolated from surgical intensive care units and to establish the prevalence of extended-spectrum beta-lactamases (ESBLs) and inducible beta-lactamases (IBLs). We also wished to determine the widespread beta-lactam substrate in ESBL-positive GNRs by two different E-test strips and to discuss the value of the routine utilization of these substrates together. METHODS: Out of 348 nosocomial gram-negative strains isolated with similar methods, 236 strains with resistance to the beta-lactam group of antimicrobials using the E-test method were included in this study. Two different strips were used for the detection of ESBLs: cefotaxime/cefotaxime + clavulanic acid (CT/CTL) and ceftazidime/ceftazidime + clavulanic acid (TZ/TZL). For IBLs, the double-disk method was used. RESULTS: The order of frequency of the strains, starting with the most frequent, was Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa. In the 236 strains, the ESBL positivity rate was found to be 19.5% with TZ/TZL and CT/CTL strips, while it was 13.2% for IBL in 348 strains. Seventy-one percent of ESBL-positive strains gave parallel results with TZ/TZL and CT/CTL. ESBL positivity with only TZ/TZL or only CT/CTL was found to be 18 and 8%. CONCLUSIONS: Our study showed that except for imipenem, amikacin and ciprofloxacin, there was a high resistance to other antimicrobials, and multiresistance rates were increased in the strains in which ESBLs and IBLs were detected. In particular, the increasing prevalence of ESBLs in K. pneumoniae and IBLs in P. aeruginosa emphasizes the importance of the problem of infection control and antibiotic administration policies. Although it was seen that the prevalence of substrate templates in the detection of ESBL positivity was similar, we think that it is more useful to use two different strips together to obtain precise results.

Anti-Bacterial Agents↗

Prevalence of extended-spectrum beta-lactamases in Enterobacteriaceae, Pseudomonas and Stenotrophomonas as determined by the VITEK 2 and E test systems in a Kuwait teaching hospital.

OBJECTIVE: To determine the prevalence of extended-spectrum beta-lactamase (ESBL)-producing members of the Enterobacteriaceae using VITEK 2 and E test systems. MATERIALS AND METHODS: A total of 3,592 consecutive gram-negative isolates (single isolate per patient) of the family of Enterobacteriaceae and Pseudomonas adjudged to be clinically relevant to the patient's infection were studied for ESBL production over a period of 1 year at Mubarak Al-Kabeer Hospital, Kuwait. Two methods were used: the automated VITEK 2 system and E test ESBL, a manually manipulated plastic strip containing various gradients of beta-lactam antibiotics. These tests and interpretative criteria for the results were performed according to the manufacturer's instructions. RESULTS: Of the 3,592 bacterial isolates, 264 (7.5%) and 185 (5.2%) were positive for ESBL production by the VITEK 2 and E test, respectively. All the ESBL-producing Pseudomonas aeruginosa identified by VITEK 2 gave indeterminate results by E test. Prevalent ESBL producers, identified by the VITEK 2 versus E test, respectively, were: Citrobacter spp. (15 vs. 3.2%), K. pneumoniae (12.2 vs. 11.4%), Enterobacter spp. (12 vs. 3%), E. coli (6.5 vs. 5.6%), P. aeruginosa (6.5 vs. 0%) and Morganella spp. (2 vs. 1%). The most common infection associated with ESBL-producing pathogens was urinary tract infection (68.2%), followed by wound infection (14.4%) and bloodstream infection (6.1%). CONCLUSION: The result of this study showed a relatively high prevalence of clinically significant ESBL producers among the Enterobacteriaceae and Pseudomonas spp. at our teaching hospital. The VITEK 2 identified a higher prevalence of ESBL strains than the E test.

Anti-Bacterial Agents↗

Prevalence and antimicrobial susceptibility of extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae in a general hospital.

BACKGROUND: The prevalence of extended-spectrum beta-lactamases (ESBLs) varies between countries and institutions. We studied the prevalence of ESBL among clinical isolates of Escherichia coli and Klebsiella pneumoniae and analyzed patterns of susceptibilities to different antimicrobial agents in a general hospital in Saudi Arabia over a 15-month period. METHODS: A total of 2455 clinical isolates of E. coli and K. pneumoniae were tested for ESBL production by double-disk diffusion. The minimum inhibitory concentration to imipenem, meropenem, piperacillin-tazobactam, cefepime, ciprofloxacin, gentamicin and amikacin were determined by the agar dilution method. RESULTS: Of the 2455 isolates of E. coli and K. pneumoniae tested, 268 (11%) produced ESBL. The ESBL phenotype was detected in 10.3% of 1674 E. coli isolates and 12.2% of 781 K. pneumoniae isolates. The majority of these isolates were from urine (57.5%) and wounds (17%). Only 7% of the blood culture isolates were ESBL-producing. Overall, carbapenems (imipenem and meropenem) had good activity against the ESBL-producing isolates tested (over 92% of isolates were susceptible). There was no difference in the activity of imipenem and meropenem against the ESBL-producing E. coli or K. pneumoniae. Over 66% of the isolates were susceptible to piperacillin-tazobactam. Susceptibilities of the isolates to amikacin varied, ranging from 72.8% for E. coli to 62% for K. pneumoniae. Gentamicin, ciprofloxacin and cefepime were active against 58.6%, 55% and 22.8% of the isolates, respectively. CONCLUSION: Our findings demonstrate the increasing incidence of infection with ESBL-producing bacteria, and the high rates of antimicrobial resistance encountered among them. Clinicians should be familiar with the clinical importance of these enzymes and potential strategies for dealing with them.

Anti-Bacterial Agents↗

International prospective study of Klebsiella pneumoniae bacteremia: implications of extended-spectrum beta-lactamase production in nosocomial Infections.

BACKGROUND: Commonly encountered nosocomially acquired gram-negative bacteria, especially Klebsiella pneumoniae, produce extended-spectrum beta-lactamases (ESBLs) as an antibiotic resistance mechanism. OBJECTIVE: To determine whether microbiology laboratories should report the presence of ESBLs and to establish the infection-control implications of ESBL-producing organisms. DESIGN: Prospective observational study. SETTING: 12 hospitals in South Africa, Taiwan, Australia, Argentina, the United States, Belgium, and Turkey. PATIENTS: 440 patients with 455 consecutive episodes of K. pneumoniae bacteremia between 1 January 1996 and 31 December 1997; of these, 253 episodes were nosocomially acquired. MEASUREMENTS: The K. pneumoniae isolates were examined for the presence of ESBLs. Pulsed-field gel electrophoresis was used to analyze the molecular epidemiology of nosocomial bacteremia with ESBL-producing K. pneumoniae. RESULTS: Overall, 30.8% (78 of 253) episodes of nosocomial bacteremia and 43.5% (30 of 69) episodes acquired in intensive care units were due to ESBL-producing organisms. After adjustment for potentially confounding variables, previous administration of beta-lactam antibiotics containing an oxyimino group (cefuroxime, cefotaxime, ceftriaxone, ceftazidime, or aztreonam) was associated with bacteremia due to ESBL-producing strains (risk ratio, 3.9 [95% CI, 1.1 to 13.8]). In 7 of 10 hospitals with more than 1 ESBL-producing isolate, multiple strains with the same genotypic pattern were observed, indicating patient-to-patient spread of the organism. CONCLUSIONS: Production of ESBLs by Klebsiella pneumoniae is a widespread nosocomial problem. Appropriate infection control and antibiotic management strategies are needed to stem the spread of this emerging form of resistance.

Bacteremia↗

[Drug resistance and genotyping of Klebsiella pneumoniae in lower respiratory tract infection].

OBJECTIVE: To understand the current drug resistance of Klebsiella pneumoniae in lower respiratory tract infection and the prevalence trend of extended spectrum beta-lactamase (ESBLs)-producing Klebsiella pneumoniae. METHODS: Drug resistance in 86 strains of Klebsiella pneumoniae and ESBLs-producing strains were genotyped by randomly amplified DNA polymorphisms (RAPD). RESULTS: ESBLs-producing strains accounted for 20.9% of the 86 strains, and 83.3% of the ESBLs-producing strains were from respiratory ICU. The drug resistance rate of ESBLs positive bacteria was much higher than that of ESBLs negative bacteria. ESBLs positive bacteria were resistant to both the quinolones and a minoglylosides. Eighteen ESBLs-producing strains were divided into 12 types. 3 strains had the same profile (genotype A), and 5 strains had an identical profile (genotype B), but other strains had different profiles. CONCLUSIONS: Chromosomal DNA analysis proved that certain types of ESBLs-producing Klebsiella pneumoniae were epidemic in respiratory ICU. RAPD was an economic, simple but reliable method in epidemic study.

Adolescent↗

[Drug-resistance analysis of Gram-negative bacilli producing extended-spectrum beta-lactamases in lower respiratory tract infection].

OBJECTIVE: To analyze occurrence of Gram-negative bacteria producing extended-spectrum beta-lactamase (ESBL) in patients with lower respiratory tract infection and assess their drug sensitivity to provide evidence for clinical management of lower respiratory tract infection with drug-resistant ESBL-producing bacteria. METHODS: A total of 312 Gram-negative bacterial strains were isolated from patients with lower respiratory tract infection and identified for the presence of ESBL by Vitek ESBL detection test and double-disk synergy tests as well as standard disk diffusion test. The effects of 10 antibacterial drugs on ESBL-producing strains were compared. RESULTS: A rate of 20.1% of the isolated strains were identified to produce ESBLs, including Klebsiella Pneumoniae, Escherichia coli, Enterobacter cloacae, Pseudomonas aeruginasa and Klebsiella oxytocla (accounting for 30.1%, 29.1%, 27.0%, 11.3%, and 2.5%, respectively, of the ESBL-producing strains). Imipenem possessed the most powerful antibacterial activity against ESBL-producing strains. CONCLUSIONS: Attention should be given to lower respiratory tract infections with ESBL-producing Gram-negative bacilli, and imipenem can be the primary choice for fighting such infections.

Drug Resistance, Bacterial↗

Extended-spectrum beta-lactamases in urinary isolates of Escherichia coli, Klebsiella pneumoniae and other gram-negative bacteria in a hospital in Eastern Province, Saudi Arabia.

OBJECTIVE: To evaluate the prevalence and antimicrobial susceptibility of extended spectrum beta-lactamase (ESBL) producing gram-negative organisms isolated from patients with urinary tract infection (UTI). METHODS: We carried out this study at Almana General Hospital, Eastern Province, Kingdom of Saudi Arabia, during the period August 2003 to October 2004. We studied urinary isolates of Escherichia coli (E. coli), Klebsiella pneumoniae (K. pneumoniae), Enterobacter spp and Pseudomonas aeruginosa (P. aeruginosa) for ESBL production and antimicrobial susceptibility. RESULTS: We studied a total of 2302 urinary gram-negative isolates for the presence of ceftazidime resistance and ESBL. The isolates included E. coli (1238), K. pneumoniae (522), Enterobacter spp (138) and P. aeruginosa (404). Of the 2302 isolates, 232 (10%) were ceftazidime resistant and 204 (8.9%) were ESBL producers. We detected ESBL in 119 (9.6%) E. coli, 59 (11.3%) K. pneumoniae, 14 (10.14%) Enterobacter species and 12 (2.97%) P. aeruginosa isolates. The ESBL-producing strains were most commonly isolated from patients with indwelling Foley s catheter [131 (64.2%)] and those in the long-term care ward [90 (44.2%)]. Only 26 (12.7%) ESBL-producing isolates were from outpatients. More than 89% of the ESBL producers were susceptible to imipenem and meropenem. Amikacin and piperacillin/tazobactam were active against 68% and 45% of the isolates. Susceptibility to gentamicin and ciprofloxacin was 22.5% and 14%. The least active antibiotic was cefepime (11.8%). CONCLUSION: This study shows the presence of ESBL producers in uropathogens from both inpatients and outpatients and demonstrates their high resistance to various classes of antimicrobial agents.

Anti-Bacterial Agents↗

Epidemiology of extended-spectrum beta-lactamase producing gram-negative bacilli at Siriraj Hospital, Thailand, 2003.

A cross-sectional study was conducted from August to September, 2003 to determine the prevalence and risk factors in acquiring extended-spectrum beta-lactamase (ESBL) producing gram-negative bacilli (GNB) in patients admitted to Siriraj Hospital and the outcomes of these infections. Of 346 isolates of gram-negative bacteria in 249 patients, 102 isolates from 87 patients were colonization only, but 244 isolates from 162 patients were infections. The common GNB were Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Acinetobacter baumannii and Enterobacter cloacae. The overall prevalence of ESBL producers was 30.1%. K. pneumoniae had a very high prevalence of ESBL producers (56.9%). The urinary tract was the most common site for ESBL- producing GNB infections. Nosocomial infections, duration from admission to infection, peripheral line, urinary catheterization, nasogastric tube insertion and previous use of beta-lactams, cephalosporins and fluoroquinolones were associated with acquiring ESBL-producing GNB infections. ESBL-producing GNB were significantly more resistant to antimicrobial agents. More than 80% of ESBL-producing GNB were susceptible to carbapenems. Mortality in patients infected with ESBL-producing GNB (41.3%) was significantly higher than those infected with non- ESBL-producing GNB (19.8%).

Carbapenems↗

The prevalence of extended-spectrum beta lactamase-producing Escherichia coli and Klebsiella pneumoniae in clinical isolates and risk factors.

OBJECTIVE: To determine the prevalence of extended-spectrum beta-lactamase (ESBL) producing Klebsiella pneumoniae (K. pneumoniae) and Escherichia coli (E. coli), risk factors of ESBL-producing strains and antimicrobial susceptibility pattern of ESBL-producing and non producing strains. METHODS: The study took place at the Faculty of Medicine, Osmangazi University, Eskisehir, Turkey from March to November 2002. We evaluated 100 K. pneumoniae and 100 E. coli strains isolated from various clinical specimens, as well as the patients from whom these strains were isolated. The double-disk synergy test was performed on the isolates for the detection of ESBL. We visited the patients with a growth of E. coli or K. pneumoniae or both from their clinical specimens in their wards if they were hospitalized, while the outpatients with a growth of these microorganisms were evaluated from their hospital records. RESULTS: The prevalence of ESBL-producing K. pneumoniae was 47% and E. coli was found as 12%. The ESBL-producing isolate rates were 50% (14/28) in intensive care units, 36.1% (35/97) in wards and 13.3% (10/75) in outpatients. Foley catheter (p<0.001), intravenous catheter (p<0.001), central venous catheter (p=0.002), intubation (p<0.001), surgery (p<0.001) and mechanical ventilation (p=0.002) were found as the risk factors for the acquisition of E. coli and K. pneumoniae with ESBLs. CONCLUSION: In our study, the prevalence of ESBL-producing isolates was high. The results of the study suggest that an antimicrobial policy and early removal of interventional apparatus be of importance for the control of ESBL-producing K. pneumoniae and E. coli.

Adult↗

Prevalence of beta-lactams resistance among Escherichia coli clinical isolates from a hospital in Algiers.

A high prevalence of beta-lactams resistance among Enterobacteriaceae have been reported worldwide; however, there are not sufficient data on this issue in Algeria. beta-Lactams susceptibility of 203 Escherichia coli clinical isolates was determined by agar diffusion method, and production of extended-spectrum beta-lactamases (ESBL) was screened by double-disk synergy test. This analysis showed five well-defined phenotypes: 1) 62 isolates (30.5%) were susceptible to all beta-lactams; 2) 135 isolates (66.5%) presented a broad-spectrum beta-lactamases phenotype (BSBL); 3) three isolates (1.5%) were defined as producing ESBLs; 4) two isolates (1%) were AmpC cephalosporinase producers; and 5) one isolate (0.5%) presented a phenotype of cell-decreased permeability to beta-lactams. Isoelectric focusing revealed beta-lactamases with isolectric points of 5.4 or 7.6 for isolates with BSBL phenotype; approximately 9.0 for two ESBL isolates; 5.4, 7.6 and approximately 9.0 for the remaining ESBL isolate; and 5.4 and approximately 9.0 for the AmpC isolates. The cefotaxime hydrolysis corresponds to the basic bands with an isoelectric point of approximately 9.0. Conjugation assay showed transfer of penicillinase and AmpC resistance phenotypes and their corresponding beta-lactamases to recipient E. coli BM21 in association with plasmids of 71.4 kb for the AmpC isolates and from 40-56 kb for penicillinase isolates. This result showed that the AmpC phenotype is plasmid mediated. ESBL isolates were found not to transfer their resistance through conjugation experiment. Polymerase chain reaction (PCR) experiments using primers specific to blaTEM, blaAmpC and blaCTX-M genes showed specific amplification with blaCTX-M primer for two ESBL isolates; blaTEM and blaCTX-M for the remaining ESBL isolate; and blaTEM and blaAmpC for the AmpC isolates and their corresponding transconjugants. The study showed a high rate of isolates producing penicillinase, and low frequencies of AmpC and ESBL phenotypes. AmpC beta-lactamases were plasmid mediated, and ESBLs belong to the CTM-M type.

Algeria↗

Occurrence of extended-spectrum beta-lactamases in members of the family Enterobacteriaceae in Italy: implications for resistance to beta-lactams and other antimicrobial drugs.

An Italian nationwide survey was carried out to assess the prevalences and the antimicrobial susceptibilities of members of the family Enterobacteriaceae producing extended-spectrum beta-lactamases (ESBLs). Over a 6-month period, 8,015 isolates were obtained from hospitalized patients and screened for resistance to extended-spectrum cephalosporins and monobactams. On the basis of a synergistic effect between clavulanate and selected beta-lactams (ceftazidime, aztreonam, cefotaxime, cefepime, and ceftriaxone), 509 isolates were found to be ESBL positive (6.3%). Colony blot hybridization with bla(TEM) and bla(SHV) DNA probes allowed one to distinguish four different genotypes: TEM-positive, SHV-positive, TEM- and SHV-positive, and non-TEM, non-SHV ESBL types. MICs for each isolate (E-test) were obtained for widely used beta-lactams, combinations of beta-lactams with beta-lactamase inhibitors, aminoglycosides, and fluoroquinolones. Among ESBL-positive strains, Klebsiella pneumoniae, Proteus mirabilis, and Escherichia coli accounted for 73.6% of isolates. Overall, TEM-type ESBLs were more prevalent than SHV-type enzymes (234 versus 173), whereas the prevalence of strains producing both TEM- and SHV-type ESBLs was similar to that of isolates producing non-TEM, non-SHV enzymes (55 and 38, respectively). In vitro, all but one of the ESBL-producing isolates remained susceptible to imipenem. Susceptibility to other drugs varied: piperacillin-tazobactam, 91%; amoxicillin-clavulanic acid, 85%; cefoxitin, 78%; amikacin, 76%; ampicillin-sulbactam, 61%; ciprofloxacin, 58%; and gentamicin, 56%. Associated resistance to aminoglycosides and ciprofloxacin was observed most frequently among TEM-positive strains. Since therapeutic options for multiresistant Enterobacteriaceae are limited, combinations of beta-lactams and beta-lactamase inhibitors appear to represent an important alternative for treating infections caused by ESBL-producing ENTEROBACTERIACEAE:

Aminoglycosides↗

Genomic Diversity and Extended-Spectrum &#x3b2;-Lactamase Gene Contexts of Community Resident-Carried Escherichia coli in Ecuador.

Community carriage of extended-spectrum &#x3b2;-lactamase (ESBL)-producing Escherichia coli represents an important reservoir of antimicrobial resistance. However, the genomic diversity and population structure of ESBL-producing E. coli circulating in community settings remain poorly characterized. This study aimed to characterize ESBL-producing E. coli isolated from fecal samples of residents in Ecuador, with an emphasis on the diversity and genomic context of ESBL genes. ESBL-producing E. coli was isolated from fecal samples obtained from 55 residents using MacConkey agar supplemented with cefotaxime. Whole-genome sequencing of the isolates was performed using a hybrid approach combining long- and short-read platforms. Plasmids and &#x3b2;-lactamase genes were identified using DFAST and PlasmidFinder. Bacterial identification and antimicrobial susceptibility testing were conducted by MALDI-TOF MS and the broth microdilution method, respectively. ESBL-producing E. coli were isolated from 35 of 55 fecal samples (63.6%). Complete circular genomes were obtained from 31 isolates. All isolates harbored bla CTX-M genes, predominantly belonging to the bla CTX-M-1 group, whereas 65.7% carried bla TEM, mainly bla TEM-1, and related variants. Although &#x3b2;-lactamase genes were predominantly plasmid-borne, chromosomal integration was detected in 40% of the isolates. Notably, 87.5% of the isolates harbored IncF plasmids with multiple replicons. Conserved IS26-flanked transposons carrying bla CTX-M and bla TEM were frequently identified in the plasmids. Phylogenetic analysis revealed substantial genomic diversity across seven phylogroups, together with closely related isolates detected within and between households. These findings provide high-resolution genomic insights into the ESBL determinants circulating in community residents and reveal region-specific patterns of ESBL genomic diversity.

CTX-M &#x3b2;-lactamases↗

Impact of inadequate initial antimicrobial therapy on mortality in infections due to extended-spectrum beta-lactamase-producing enterobacteriaceae: variability by site of infection.

BACKGROUND: Infections due to extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella species (ESBL-EK) have increased markedly in recent years. Risk factors for mortality among ESBL-EK infections have not been studied. METHODS: This retrospective cohort study was conducted in a 625-bed tertiary care medical center and a 344-bed urban community hospital to determine whether inadequate initial antimicrobial therapy (IIAT) (>48 hours between the time a culture was obtained and initiation of an agent to which the infecting organism was susceptible) is associated with mortality in ESBL-EK infections. All hospitalized patients with an ESBL-EK infection between June 1, 1997, and December 31, 2002, were eligible for inclusion. Subsequently, we conducted a nested case-control study to identify risk factors for IIAT. RESULTS: Of 187 subjects, 32 (17.1%) died while in the hospital. Clinical site of infection was a significant effect modifier in the association between IIAT and mortality. The presence of IIAT was an independent risk factor for mortality, but only for nonurinary ESBL-EK infections (adjusted odds ratio [95% confidence interval], 10.04 [1.90-52.96]). Independent risk factors for IIAT were (1) infection with a multidrug-resistant ESBL-EK (ie, resistant to sulfamethoxazole-trimethoprim, aminoglycosides, and quinolones) (14.58 [1.91-111.36]) and (2) health care-acquired ESBL-EK infection (4.32 [1.49-12.54]). CONCLUSIONS: Inadequate initial antimicrobial therapy is an independent risk factor for mortality in ESBL-EK infections, but only among nonurinary infections. Multidrug resistance was a strong risk factor for IIAT.

Aged↗

Detection of genes coding for extended-spectrum SHV beta-lactamases in clinical isolates by a molecular genetic method, and comparison with the E test.

A highly sensitive and specific method, termed PCR/NheI, for the detection of genes coding for SHV extended-spectrum beta-lactamases (ESBL) in clinical isolates is presented. It is based on polymerase chain reaction (PCR) amplification of the blaSHV genes, followed by restriction with NheI. Due to the glycine (positive 238) (SHV-non-ESBL)-->serine (position 238) (SHV-ESBL) mutation, only PCR fragments from the genes coding for SHV-ESBLs were cleaved. A commercially available test for ESBLs, the E test ESBL, identified 52% of our 29 clinical isolates carrying blaSHV-ESBL genes as ESBL producers.

Base Sequence↗

Use of boronic acid disk methods to detect the combined expression of plasmid-mediated AmpC beta-lactamases and extended-spectrum beta-lactamases in clinical isolates of Klebsiella spp., Salmonella spp., and Proteus mirabilis.

A study using boronic acid (BA), an AmpC enzyme inhibitor, was designed to detect the combined expression of plasmid-mediated AmpC beta-lactamases (pAmpCs) and extended-spectrum beta-lactamases (ESBLs) in bacterial isolates naturally lacking chromosomal ampC genes. A total of 122 Klebsiella spp., Salmonella spp., and Proteus mirabilis isolates producing or nonproducing pAmpCs and/or ESBLs were analyzed. Detection of genes encoding ESBLs and AmpCs was confirmed by polymerase chain reaction (PCR) followed by sequencing of PCR products. A > or = 5-mm increase in zone diameter for i) cefoxitin (FOX) and/or cefotetan (CTT) containing BA versus FOX and/or CTT alone was considered positive for AmpC; ii) ceftazidime (CAZ)-clavulanate (CA) and/or cefotaxime (CTX)-CA tested in combination with BA versus CAZ and/or CTX containing BA was considered positive for ESBL. The disk tests of FOX and/or CTT alone and with BA detected 98.4% of organisms producing pAmpCs. All of the 21 pAmpC and ESBL coproducers were accurately detected ESBL by the disk tests of CTX-CA and/or CAZ-CA containing BA and CTX and/or CAZ containing BA. In conclusion, The BA disk test using Clinical and Laboratory Standards Institute methodology is simple and very efficient method to detect pAmpC and ESBL in organisms naturally lacking chromosomal AmpC enzymes. In particular, the method accurately detects the isolates that harbor both AmpCs and ESBLs.

Bacterial Proteins↗

Prevalence of extended-spectrum beta-lactamase-positive Klebsiella pneumoniae isolates in the Czech Republic.

This study focused on the prevalence and molecular biology of extended-spectrum beta-lactamase (ESBL)-positive Klebsiella pneumoniae isolates collected in the Czech Republic. Clinical material from patients hospitalised in 16 Czech hospitals in September 2004 was used to isolate K. pneumoniae strains. Strains were identified by standard identification procedures. Susceptibility of the strains to antibiotics was tested using a microdilution method. The double-disk synergy test and combination disk method were used to determine ESBL production. Molecular biology characteristics of ESBL-positive isolates were determined using genomic DNA isolation, XbaI restriction digestion and pulsed-field gel electrophoresis differentiation. The acquired restriction maps of individual isolates were compared using GelCompar II software and their relationships were determined. During the 3-week period, 483 K. pneumoniae strains causing clinically detectable diseases were isolated. Of these, 117 (24.2%) were determined to be ESBL-positive. The prevalence of ESBL-positive isolates was 38.9% in Intensive Care Units (ICUs) and 13.1% in standard wards. More than 50% of ESBL-positive isolates were treated effectively only with meropenem (98%), cefoperazone/sulbactam (61%) and amikacin (54%). Conversely, ESBL-negative isolates showed high susceptibility to all tested antibiotics (76-99%). Molecular biology analysis identified 18 clonal types containing two to six identical isolates. Seventeen clones usually contained isolates from only one hospital; isolates from two hospitals were identified only in one clone. Based on the abovementioned results, the prevalence of ESBL-positive K. pneumoniae isolates in the Czech Republic can be perceived as relatively high, especially in ICUs. Extensive spread of 'epidemic clones' within Czech hospitals and, to a limited extent, between them can be demonstrated.

Czech Republic↗

Characteristics, epidemiology and clinical importance of emerging strains of Gram-negative bacilli producing extended-spectrum beta-lactamases.

Beta-lactam antimicrobial agents represent the most common treatment for bacterial infections and continue to be the leading cause of resistance to beta-lactam antibiotics among Gram-negative bacteria worldwide. The persistent exposure of bacterial strains to a multitude of beta-lactams has induced dynamic and continuous production and mutation of beta-lactamases in these bacteria, expanding their activity even against the newly developed beta-lactam antibiotics. These enzymes are known as extended-spectrum beta-lactamases (ESBLs). The majority of ESBLs are derived from the widespread broad-spectrum beta-lactamases TEM-1 and SHV-1. There are also new families of ESBLs, including the CTX-M and OXA-type enzymes as well as novel unrelated beta-lactamases. In recent years, there has been an increased incidence and prevalence of ESBLs. ESBLs are mainly found in strains of Escherichia coli and Klebsiella pneumoniae but have also been reported in other Enterobacteriaceae strains and Pseudomonas aeruginosa. Infections with ESBL-producing bacterial strains are encountered singly or in outbreaks, especially in critical care units in hospitals, resulting in increasing cost of treatment and prolonged hospital stays. Not only may nursing home patients be an important reservoir of ESBL-containing multiple antibiotic-resistant organisms, but ambulatory patients with chronic conditions may also harbor ESBL-producing organisms.

Anti-Bacterial Agents↗