scholarly journals Molecular characterization of extended-spectrum beta-lactamases in Enterobacteriaceae from patients of two hospitals in Saxony, Germany

2007 ◽  
Vol 56 (2) ◽  
pp. 241-249 ◽  
Author(s):  
Joachim Schmitt ◽  
Enno Jacobs ◽  
Herbert Schmidt

Between January and September 2003, 39 isolates of the family Enterobacteriaceae with phenotypically positive Vitek 1 extended-spectrum beta-lactamase (ESBL) test results were collected, originating from patients of two hospitals in Saxony, Germany. Plasmid DNA was isolated and screened by PCR for the presence of genes encoding beta-lactamases of SHV, TEM and CTX-M types. To differentiate ESBL and non-ESBL among SHV and TEM genes, detailed analysis of PCR products was performed. Twenty-four strains carried SHV-2, SHV-5 or SHV-12 genes. In a further 11 strains a CTX-M gene was detected. The CTX-M genes could be affiliated to the CTX-M-1 and CTX-M-9 cluster by RFLP analysis. In the case of four Klebsiella oxytoca isolates, hyperproduction of the chromosomal beta-lactamase K1 was inferred, because genes of the above-mentioned types were not detected. The strains contained plasmid DNA between 45 and 160 kb in size. Common plasmid restriction patterns among SHV-5 producers provided evidence of horizontal spread. Twenty strains had a MIC for cefotaxime of ⩽4 mg l−1, 18 strains had the same MIC for ceftazidime, and nine strains had this MIC of >4 mg l−1 for both antibiotics. The ESBL phenotypes often coincided with ciprofloxacin or gentamicin resistance.

1996 ◽  
Vol 40 (4) ◽  
pp. 1027-1029 ◽  
Author(s):  
M J Soilleux ◽  
A M Morand ◽  
G J Arlet ◽  
M R Scavizzi ◽  
R Labia

Crude extracts from 115 extended-spectrum beta-lactamase-producing Klebsiella pneumoniae isolates were analyzed biochemically. The TEM-3 type was encountered 108 times, SHV types were encountered 7 times, and the TEM-26 type was encountered only once. For the last one, the gene was identified; an adenine was detected at position 925, as in blaTEM-26B not in blaTEM-26.


Medicina ◽  
2007 ◽  
Vol 43 (10) ◽  
pp. 778
Author(s):  
Astra Vitkauskienė ◽  
Agnė Giedraitienė ◽  
Vytis Dudzevičius ◽  
Raimundas Sakalauskas

Aim of the study. To evaluate relationship between isolation of extended spectrum beta-lactamaseproducing Klebsiella pneumoniae strains and course of hospital-acquired pneumonia. Materials and methods. K. pneumoniae strains isolated from bronchial secretions or bronchoalveolar lavage fluid samples of patients hospitalized at an intensive care unit of Kaunas University of Medicine Hospital were analyzed. By means of synergistic two-antibiotics disc method, K. pneumoniae strains producing extended spectrum beta-lactamases were selected for further analysis using E-test (AB Biodisk, Solna, Sweden). Hospitalacquired pneumonia was diagnosed based on standard criteria for the diagnosis of pneumonia if signs of pneumonia occurred after 48 hours following admission. Late-onset hospital-acquired pneumonia was considered if these signs of pneumonia occurred on fifth day of hospitalization or later. Results. Total of 45 strains of K. pneumoniae were isolated during the study period; 18 isolated strains produced ESBL. Thirty-two patients investigated have developed hospital-acquired pneumonia, 20 of which were cases of late-onset hospital-acquired pneumonia. Thirteen cases of K. pneumoniae isolation were classified as airway colonization. Extended spectrum beta-lactamase-producing K. pneumoniae strains were more frequently isolated from patients with hospital-acquired pneumonia (88.9%, n=16 and 11.1%, n=2, P<0.05) in comparison with non-producing strains. Extended spectrum beta-lactamase-producing strains were more prevalent in late-onset pneumonia group (93.8%, n=15) than in early-onset group (6.2%, n=1, P<0.001). Conclusions. Extended spectrum beta-lactamase-producing K. pneumoniae strains were more frequently isolated from patients with hospital-acquired pneumonia as compared to colonized patients. Extended spectrum beta-lactamase-producing K. pneumoniae strains were more frequently isolated from patients with late-onset hospital-acquired pneumonia.


2008 ◽  
Vol 74 (23) ◽  
pp. 7439-7441 ◽  
Author(s):  
Patricia Poeta ◽  
Hajer Radhouani ◽  
Gilberto Igrejas ◽  
Alexandre Gonçalves ◽  
Carlos Carvalho ◽  
...  

ABSTRACT Escherichia coli isolates containing the following extended-spectrum beta-lactamases have been detected in 11 of 57 fecal samples (19.3%) in Berlengas Island seagulls: TEM-52 (eight isolates), CTX-M-1 (one isolate), CTX-M-14a (one isolate), and CTX-M-32 (one isolate). Most of the extended-spectrum beta-lactamase-positive isolates harbored class 1 or class 2 integrons, which included different antibiotic resistance gene cassettes.


2005 ◽  
Vol 49 (11) ◽  
pp. 4716-4720 ◽  
Author(s):  
Dóra Szabó ◽  
Melissa A. Melan ◽  
Andrea M. Hujer ◽  
Robert A. Bonomo ◽  
Kristine M. Hujer ◽  
...  

ABSTRACT Bacteria that simultaneously produce multiple extended-spectrum beta-lactamases are frequently isolated. We report an Enterobacter cloacae isolate, ES24, producing four different beta-lactamases (AmpC type beta-lactamase, TEM-1, SHV-7, and a novel extended-spectrum beta-lactamase, SHV-30). Direct sequencing of bla SHV gene products gave a “double peak” at position 703, suggesting the presence of more than one allele. Using fluorescence resonance energy transfer real-time PCR to detect single-nucleotide polymorphisms, we were able to distinguish two different bla SHV genes in a single isolate. This may prove to be a useful technique in surveys of beta-lactamase production in contemporary clinical isolates.


2005 ◽  
Vol 49 (11) ◽  
pp. 4745-4750 ◽  
Author(s):  
Inna Chmelnitsky ◽  
Yehuda Carmeli ◽  
Azita Leavitt ◽  
Mitchell J. Schwaber ◽  
Shiri Navon-Venezia

ABSTRACT The rate of occurrence of the extended-spectrum beta-lactamase (ESBL)-producing phenotype among Escherichia coli isolates in Tel Aviv is 12% (22). The aim of this study was to understand the molecular epidemiology of E. coli ESBL producers and to identify the ESBL genes carried by them. We studied 20 single-patient ESBL-producing E. coli clinical isolates. They comprised 11 distinct nonrelated pulsed-field gel electrophoresis (PFGE) genotypes: six isolates belonged to the same PFGE clone, four other clones included two isolates each, and six unrelated clones included only one isolate. All isolates produced various beta-lactamases with pIs ranging from 5.2 to 8.2, varying within similar PFGE clones. The most prevalent ESBL gene was bla CTX-M; 16 isolates carried bla CTX-M-2 and three carried a new ESBL gene designated bla CTX-M-39. Three strains carried bla SHV (two bla SHV-12 and one bla SHV-5), and two strains carried inhibitor-resistant ESBL genes, bla TEM-33 and bla TEM-30; 18 strains carried bla TEM-1 and eight strains carried bla OXA-2. Plasmid mapping and Southern blot analysis with a CTX-M-2 probe demonstrated that bla CTX-M-2 is plasmid borne. The wide dissemination of ESBLs among E. coli isolates in our institution is partly related to clonal spread, but more notably to various plasmid-associated ESBL genes, occurring in multiple clones, wherein the CTX-M gene family appears almost uniformly. We report here a new CTX-M gene, designated bla CTX-M-39, which revealed 99% homology with bla CTX-M-26, with a substitution of arginine for glutamine at position 225.


1996 ◽  
Vol 40 (3) ◽  
pp. 616-620 ◽  
Author(s):  
A Bauernfeind ◽  
I Stemplinger ◽  
R Jungwirth ◽  
P Mangold ◽  
S Amann ◽  
...  

Plasmidic extended-spectrum beta-lactamases of Ambler class A are mostly inactive against ceftibuten. Salmonella typhimurium JMC isolated in Argentina harbors a bla gene located on a plasmid (pMVP-5) which confers transferable resistance to oxyiminocephalosporins, aztreonam, and ceftibuten. The beta-lactamase PER-2 (formerly ceftibutenase-1; CTI-1) is highly susceptible to inhibition by clavulanate and is located at a pI of 5.4 after isoelectric focusing. The blaPER-2 gene was cloned and sequenced. The nucleotide sequence of a 2.2-kb insert in vector pBluescript includes an open reading frame of 927 bp. Comparison of the deduced amino acid sequence of PER-2 with those of other beta-lactamases indicates that PER-2 is not closely related to TEM or SHV enzymes (25 to 26% homology). PER-2 is most closely related to PER-1 (86.4% homology), an Ambler class A enzyme first detected in Pseudomonas aeruginosa. An enzyme with an amino acid sequence identical to that of PER-1, meanwhile, was found in various members of the family Enterobacteriaceae isolated from patients in Turkey. Our data indicate that PER-2 and PER-1 represent a new group of Ambler class A extended-spectrum beta-lactamases. PER-2 so far has been detected only in pathogens (S. typhimurium, Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis) isolated from patients in South America, while the incidence of PER-1-producing strains so far has been restricted to Turkey, where it occurs both in members of the family Enterobacteriaceae and in P. aeruginosa.


2014 ◽  
Vol 8 (03) ◽  
pp. 282-288 ◽  
Author(s):  
Hoda Hassan ◽  
Baha Abdalhamid

Introduction: The aim of this study was to determine the prevalence of extended-spectrum beta-lactamase (ESBL) producing Escherichia coli (E. coli), Klebsiella pneumoniae (K. pneumoniae), and Proteus mirabilis (P. mirabilis). In addition, different methods for detection of these enzymes, including the newly introduced CHROMagar ESBL, were evaluated. Methodology: A total of 382 Enterobacteriaceae clinical isolates were obtained from King Fahad Specialist Hospital – Dammam, during 2011 and screened for production of ESBL using advanced expert system of Vitek 2, CHROMagar and ESBL-E-strips. PCR assay was used to detect blaTEM, blaSHV, and blaCTX-M genes. Susceptibility to a panel of antibiotics was determined. Results: The overall proportion of ESBL-producing enterobacterial isolates was 30.6%, which was higher in E. coli (35.8%) than in K. pneumoniae (25.7%). ESBL genotypes showed remarkable increase in the CTX-M (97.4%) compared to SHV (23.1%). The predominant ESBL was CTX-M- 15 (92.1 %). No TEM ESBL was detected in this study. The Vitek2 showed the highest sensitivity (100%), and the CHROMagar had the lowest specificity (97.3%) compared to the molecular method. All isolates were susceptible to imipenem and meropenem. Conclusions: This study confirms a high level of blaCTX-M positive ESBL isolates are circulating in the Eastern Province of Saudi Arabia. The trend of a multidrug-resistant profile associated with the recovery of the blaCTX-M gene is alarming.


2012 ◽  
Vol 4 (2) ◽  
pp. 28-31 ◽  
Author(s):  
Nawshad Muhammad Wahidur Rahman ◽  
Afzalunnessa Binte Lutfor ◽  
Sanya Tahmina Jhora ◽  
Mahmuda Yasmin ◽  
Jalaluddin Ashraful Haq

A total of 200, non-duplicate ESBL producing strains (171 Escherichia coli and 29 Klebsiella spp.) from three tertiary care hospitals were detected using screening test & double disc synergy test. All isolates were screened for the detection of CTX-M type Extended spectrum Beta-lactamase (ESBL) using PCR. Among them 133 (66.5%) were positive for CTX-M type ESBLs which include 114 (66.66%) E.coli and 19 (65.51%) Klebsiella spp. This is the first report of identifying CTX-M gene in ESBL producing Escherichia coli and Klebsiella species of different hospitals.DOI: http://dx.doi.org/10.3329/bjmm.v4i2.10829 


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