Genetic background of Escherichia coli and extended-spectrum beta-lactamase type

To assess the implication of the genetic background of Escherichia coli strains in the emergence of extended-spectrum-Beta -lactamases (ESBL), 55 TEM-, 52 CTX-M-, and 22 SHV-type ESBL-producing clinical isolates involved in various extraintestinal infections or colonization were studied in terms of phylogenetic group, virulence factor (VF) content (pap, sfa/foc, hly, and aer genes), and fluoroquinolone resistance. A factorial analysis of correspondence showed that SHV type, and to a lesser extent TEM type, were preferentially observed in B2 phylogenetic group strains that exhibited numerous VFs but were fluoroquinolone-susceptible, whereas the newly emerged CTX-M type was associated with the D phylogenetic group strains that lacked VF but were fluoroquinolone-resistant. Thus, the emergence of ESBL-producing E. coli seems to be the result of complex interactions between the type of ESBL, genetic background of the strain, and selective pressures in ecologic niches.     

Distribution of extended-spectrum beta-lactamase genes using a commercial DNA micro-array system

Extended-spectrum beta-lactamase (ESBL) genes are distributed worldwide and their epidemiology is complex. Using the Check-ESBL assay, the distribution of class A ESBL genes in clinical isolates of aerobic Gram-negative bacilli from three laboratories in the East of The Netherlands was determined. Four patient categories were distinguished: (i) patients admitted to an intensive care unit (ICU); (ii) non-ICU inpatients; (iii) outpatients admitted less than a year before collection of the isolate, (<1); (iv) outpatients admitted more than one-year prior to isolate collection or who had never been hospitalized (>1). From February 2009 until March 2010, out of 491 putative ESBL-positive isolates detected by the Vitek2 or Phoenix automated sensitivity testing systems, ESBL genes were detected in 247 (50.3%) by the Check-ESBL assay. Of these, 116 were from hospitalized patients (35 ICU, 81 non-ICU) and 131 were from outpatients (43 <1, 88 >1). In all, 274 ESBL genes were identified in these 247 isolates: 153 CTX-M-1 group (predominantly in E. coli and K. pneumoniae, 70.4% and 51.6% respectively), 67 CTX-M-9 group (predominantly in E. cloacae, 57.9%), 32 SHV, 14 TEM and 8 CTX-M-2 group. ESBL-producing E. cloacae were significantly more common in hospitalized patients than in outpatients, 20.7% and 3.8% respectively (P=0.001). CTX-M-9 group ESBLs were significantly more prevalent in ICU patients (P=0.003), whereas SHV ESBLs were more common in hospitalized patients than in outpatients (P<0.001). There was no significant difference in distribution of ESBL genes between the two outpatient groups.     

Dissemination of clonally related Escherichia coli strains expressing extended-spectrum beta-lactamase CTX-M-15

We analyzed 43 CTX-M-15-producing Escherichia coli isolates and 6 plasmids encoding the blaCTX-M-15 gene from Canada, India, Kuwait, France, Switzerland, Portugal, and Spain. Most isolates belonged to phylogroups B2 (50%) and D (25%). An EC-B2 strain of clonal complex sequence type (ST) 131 was detected in all countries; other B2 isolates corresponded to ST28, ST405, ST354, and ST695 from specific areas. EC-D strains were clonally unrelated but isolates from 3 countries belonged to ST405. All CTX-M-15 plasmids corresponded to IncFII group with overrepresentation of 3 HpaI-digested plasmid DNA profiles (A, B and C; 85-120kb, similarity > or =70%). Plasmid A was detected in EC-B2 strains (ST131, ST354, or ST405), plasmid C was detected in B2 and D strains, and plasmid B was confined to worldwide-disseminated ST131. Most plasmids contained blaOXA-1, aac(6')-Ib-cr, and blaTEM-1. Worldwide dissemination of CTX-M-15 seems to be determined by clonal complexes ST131 and ST405 and multidrug-resistant IncFII plasmids.     

儿科病房137株产超广谱β-内酰胺酶菌株的分布特点

随着第三代头孢菌素的广泛应用，细菌对头孢菌素产生耐药性的流行越来越广，给临床感染性疾病的治疗造成很大的困难。超广谱β-内酰胺酶（ESBLs）主要由肠杆菌科细菌产生，大肠埃希菌和克雷伯菌为其代表菌种。收集武汉市妇女儿童保健中心近1年分离的大肠埃希菌、克雷伯菌属和肠杆菌属共543株，用纸片扩散法进行ESBLs菌株初筛和确证试验，以了解该院产ESBLs菌的流行、分布状况，现报告如下。     

An outbreak of hospital-acquired Klebsiella pneumoniae bacteraemia, including strains producing extended-spectrum beta-lactamase

This study describes the clinical outcome of an outbreak of extended-spectrum beta-lactamase producing Klebsiella pneumoniae (ESBL-KP) bacteraemia. Ninety-two episodes of hospital-acquired K. pneumoniae bacteraemia were studied, 49 ESBL-KP and 43 non-ESBL-KP, from May 1993 to June 1995. Of these, 44 (90%) episodes of ESBL-KP vs. 20 (46%) episodes of non-ESBL-KP occurred in intensive care unit (ICU) patients. The incidence of K. pneumoniae bacteraemia (mainly due to ESBL-KP) increased in the ICU during the outbreak. A significant association was found between intravascular catheter-related bacteraemia and isolation of ESBL-KP [27 (56%) in the ESBL-KP group vs. 13 (30%) in the non-ESBL-KP group;P= 0.01]. The worst prognostic features were identified as age > 65 years (P= 0.02), septic shock (P< 0.001) and secondary bacteraemia (P= 0.04). High rates of resistance to beta-lactam/beta-lactamase inhibitors observed in our ESBL-KP isolates, as well as variable activity of aminoglycosides, restricts the empirical use of these antibiotics. Carbapenems should be the treatment of choice since they are uniformly active against these strains. Our study shows that ESBL-KP bacteraemia occurring in an epidemic ICU setting is mainly catheter-related. We did not find ESBL strains to be associated with a significantly poor outcome.     

Cefepime MIC as a predictor of the extended-spectrum beta-lactamase type in Klebsiella pneumoniae, Taiwan

To guide selection of carbapenems or fourth-generation cephalosporins as therapy, 110 Klebsiella pneumoniae isolates with extended-spectrum beta-lactamases from Taiwan were characterized by phenotypic (MICs), molecular, and chemical methods. MIC patterns of ceftazidime and cefepime clearly differentiate strains treatable by cefepime and those capable of efficiently hydrolyzing available cephalosporins (CTX-M series and SHV-types). Continued use of cefepime appears to be a treatment option in cases for which MIC results are available and interpreted by the criteria presented.     

Infections due to Escherichia coli producing extended-spectrum beta-lactamase among hospitalised patients: factors influencing mortality

We performed a retrospective matched-cohort study to determine the risk factors for mortality among patients with Escherichia coli infections. From January 1996 to December 2003, 100 hospitalised patients with extended-spectrum beta-lactamase (ESBL)-producing E. coli infections were compared with patients not infected with ESBL-producing E. coli. These patients were selected according to the same site of infection and the closest date of admission. Comparison of the two groups showed that empirical antibiotic therapy was more often inadequate in patients infected with ESBL-producing E. coli (44% vs 15%; P<0.01), and that early mortality (16% vs 6%; P=0.02) and overall mortality (25% vs 11%; P=0.01) were also significantly higher in patients with ESBL-producing E. coli infections. A multivariate model identified the urinary tract focus as the only independent risk factor influencing early mortality for E. coli infections [odds ratio (OR): 0.1; 95% confidence interval (CI): 0.03-0.7; P=0.01]. All 12 patients with ESBL-producing E. coli urinary tract infections treated initially with an oxyimino-beta-lactam survived. Subsequent analysis of the factors influencing early mortality in the cohort of 130 patients with a non-urinary E. coli infection found inadequate empirical antibiotic therapy as an independent risk factor for mortality only for non-urinary E. coli infections (adjusted OR: 3.0; 95% CI: 1.0-8.6; P=0.03). The study showed that hospitalised patients with ESBL-producing E. coli infections more often receive inadequate empiric antibiotic therapy and have a higher mortality rate than those infected with non-ESBL-producing strains. The site of infection strongly influences mortality. The administration of inadequate empirical antibiotic therapy is independently associated with higher mortality only among patients with non-urinary tract infections.     

Infection control implications of extended-spectrum beta-lactamase (ESBL) production by Klebsiellae and other Gram-negative bacteria

Extended-spectrum beta-lactamase (ESBL) producing organisms have been detected in Australian hospitals over the last 10 years. They are mostly encountered in very sick patients who have been hospitalised for long periods, particularly in intensive care units (ICUs). Use of invasive medical devices and broad spectrum antibiotics predispose to colonisation and infection with these organisms.Control of epidemic or endemic ESBL producing organisms is possible using a multidisciplinary approach utilising the resources of routine clinical microbiology laboratories, molecular epidemiology laboratories and clinical infection control practitioners. Many hospitals can control ESBL producing organisms using a combination of classical infection control interventions such as contact isolation and modification of antibiotic prescribing practices, in particular restriction of use of third generation cephalosporins.     

Investigation of a nosocomial outbreak of Acinetobacter baumannii producing PER-1 extended-spectrum beta-lactamase in an intensive care unit

We investigated an outbreak of Acinetobacter baumannii in an adult intensive care unit of Kosin University Gospel Hospital in Busan, Republic of Korea. The outbreak involved 10 cases of infection by A. baumannii producing PER-1 extended-spectrum beta-lactamase over a seven-month period, and was caused by a single pulsed-field gel electrophoresis clone. The epidemic isolates were characterized by slight synergy between clavulanic acid and cefepime. Isoelectric focusing of crude bacterial extracts detected two nitrocefin-positive bands with pI values of 8.0 and 5.3. Polymerase chain reaction amplification and characterization of the amplicons by restriction analysis and direct sequencing indicated that the epidemic isolates carried a bla(PER-1) determinant. The epidemic isolates were characterized by a multidrug-resistant phenotype that remained unchanged over the outbreak, including penicillins, beta-lactam/beta-lactamase inhibitor, extended-spectrum cephalosporins and monobactams. Isolation of infected patients and appropriate carbapenem therapy were successful in ending the outbreak. Our report indicates that the bla(PER-1) resistance determinant may become an emerging therapeutic problem.     

Investigation of a nosocomial outbreak of extended-spectrum beta-lactamase VEB-1-producing isolates of Acinetobacter baumannii in a hospital setting

A nosocomial outbreak of epidemiologically related VEB-1 extended-spectrum beta-lactamase-producing isolates of Acinetobacter baumannii occurred in 33 patients in an intensive care unit. A case-control study identified previous treatment with third-generation cephalosporins as the only risk factor for A. baumannii acquisition. Rationale for antibiotic use should be strengthened.     

A case-control study of risk factors associated with rectal colonization of extended-spectrum beta-lactamase producing Klebsiella sp. in newborn infants

To determine the risk factors for rectal colonization by extended-spectrum beta-lactamase (ESBL) Klebsiella sp. in 368 newborns admitted consecutively to a neonatal intensive care unit over six months, rectal swabs were cultured on admission and weekly until discharge. Eighty infants (21.7%) had ESBL Klebsiella sp. cultured from their rectal swabs. Eighty controls were selected at random from infants with negative cultures admitted within the 14-day period prior to the detection of ESBL Klebsiella sp. in the cases. Cases had significantly lower birth weight, gestational age, earlier age of admission, longer hospital stay, and higher proportions of congenital malformations, early-onset pneumonia and respiratory distress syndrome compared with controls. Significantly more cases received mechanical ventilation, nasal continuous positive airway pressure support, total parenteral nutrition, umbilical vascular catheterization, arterial line insertion, urinary bladder catheterization, and prior treatment with antibiotics. However, stepwise logistic regression analysis showed that only two independent risk factors were significantly associated with ESBL rectal colonization: duration of hospital stay [adjusted odds ratio (OR): 1.3; 95% confidence intervals (CI): 1.2, 1.4; P<0.0001) and early-onset pneumonia (adjusted OR: 8.3; 95% CI: 1.6, 43.4; P=0.01).     

A seven-year survey of Klebsiella pneumoniae producing TEM-24 extended-spectrum beta-lactamase in Nice University Hospital (1994-2000)

The aim of this study was to investigate TEM-24-producing isolates of Klebsiella pneumoniae, and their clonal dissemination in Nice University Hospital. During the 1994-2000 period, a total of 263 non-repetitive isolates of ESBL-producing K. pneumoniae were collected. Most of these isolates were highly resistant in vitro to ceftazidime, cefotaxime and aztreonam, but susceptible to cefoxitin and imipenem. Resistance profile analysis revealed seven predominant antibiotypes (P1 to P7). Isoelectric focusing evidenced beta-lactamase activity, with a chromosomal penicillinase (pl 7.7), and one or two additional enzymes with pls ranging from 5.4 to 8.2 identified as presumed TEM-1 pl 5.4, TEM-3 pl 6.3, TEM-24 pl 6.5, SHV-3 pl 7.0, SHV-4 pl 7.8, SHV-5 pl 8.2, or other unidentified beta-lactamases. Among these K. pneumoniae, 130 isolates produced TEM-24, and 115 of them were highly resistant in vitro to quinolones (antibiotype P1). This phenotype was responsible for an outbreak in a medical intensive care unit from March to September 2000. Four isolates submitted were genetical sequenced, and shared 99.9% homology with tem-24 (GenBank no. X 65253). Pulsed-field gel electrophoresis and enterobacterial repetitive intergenic consensus polymerase chain reaction (PCR) (ERIC2-PCR) applied to 28 non-epidemic and six epidemic isolates yielded concordant results. Molecular typing revealed the persistence and dissemination of a single clone of TEM-24 producing K. pneumoniae in Nice Hospital during the seven-year study period.     

Risk factors in the acquisition of extended-spectrum beta-lactamase Klebsiella pneumoniae: a case-control study in a district teaching hospital in Taiwan

A case-control study was performed to find the risk factors in the acquisition of extended-spectrum beta-lactamase (ESBL) Klebsiella pneumoniae. From 1 May 2001 to 30 September 2001, 422 isolates ofK. pneumoniae identified by the microbiological laboratory in Hsin-Chu hospital were collected, 59 of which were ESBL-producing strains. The prevalence rate was 14% (59/422). There were 43 case patients (ESBL-producing K. pneumoniae) and 86 controls (non-ESBL-producing K. pneumoniae). Tracheostomy, insertion of a Foley catheter, endotracheal tube, nasogastric tube and central venous catheter were found to be risk factors in the acquisition of K. pneumoniae with ESBLs by univariate analysis. Tracheostomy (odds ratio, 5.13; 95% CI, 1.24-21.1;P =0.023) and ceftazidime use (odds ratio, 13.40; 95% CI, 1.21-148.85; P=0.035) remained as risk factors by multivariate analysis with logistic regression. Other anti-pseudomonal agents should be used as empirical therapy to treat possible Pseudomonas aeruginosa infection in order to reduce ceftazidime use and thereby decrease the prevalence of ESBL producing strains of Enterobacteriaceae.     

Control of Enterobacteriaceae producing extended-spectrum beta-lactamase in intensive care units: rectal screening may not be needed in non-epidemic situations.

OBJECTIVE: To evaluate the usefulness of screening cultures in the control of extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae in intensive care units (ICUs). DESIGN: A 4-year retrospective study. SETTING: Two adult ICUs of a university-affiliated public hospital in France. RESULTS: A total of 7,777 specimens were analyzed and 28 (0.97%) of 2,883 screened patients had a positive result on a screening test, among the 3,678 admitted patients. Thirteen of these 28 patients were only carriers; 4 were carriers and then were colonized or infected 2, 2, 3, and 8 days later, respectively; and 11 were colonized or infected before a screening test was positive. Cluster analysis showed that the occurrence of ESBL-producing Enterobacteriaceae cross-transmission within both ICUs was limited to 9 cases. Thus, most cases (19 of 28) were probably imported. Surveillance cultures failed to detect 9 of the 19 cases. CONCLUSION: The low prevalence of ESBL-producing Enterobacteriaceae carriers on admission (0.45%) and the relative ineffectiveness of our screening test to detect imported cases suggest that systematic detection of ESBL-producing Enterobacteriaceae in ICU patients is not cost-effective and that the use of clinical cultures may be sufficient to control ESBL-producing Enterobacteriaceae in non-epidemic situations.