Molecular Analysis of the Simultaneous Production of Two SHV-Type Extended-Spectrum Beta-Lactamases in a Clinical Isolate of Enterobacter cloacae by Using Single-Nucleotide Polymorphism Genotyping

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.

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Survey of Klebsiella pneumoniae producing extended-spectrum beta-lactamases: prevalence of TEM-3 and first identification of TEM-26 in France.

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.40.4.1027 ◽

1996 ◽

Vol 40 (4) ◽

pp. 1027-1029 ◽

Cited By ~ 17

Author(s):

M J Soilleux ◽

A M Morand ◽

G J Arlet ◽

M R Scavizzi ◽

R Labia

Keyword(s):

Klebsiella Pneumoniae ◽

Beta Lactamase ◽

Beta Lactamases ◽

Extended Spectrum Beta Lactamase ◽

Extended Spectrum ◽

Crude Extracts ◽

Extended Spectrum Beta Lactamases

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.

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

Journal of Medical Microbiology ◽

10.1099/jmm.0.46670-0 ◽

2007 ◽

Vol 56 (2) ◽

pp. 241-249 ◽

Cited By ~ 23

Author(s):

Joachim Schmitt ◽

Enno Jacobs ◽

Herbert Schmidt

Keyword(s):

Plasmid Dna ◽

Klebsiella Oxytoca ◽

Rflp Analysis ◽

Beta Lactamase ◽

M Gene ◽

Beta Lactamases ◽

Extended Spectrum Beta Lactamase ◽

Extended Spectrum ◽

Genes Encoding ◽

Extended Spectrum Beta Lactamases

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.

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Relationship between isolation of extended spectrum beta-lactamase-producing Klebsiella pneumoniae and course of hospital-acquired pneumonia

Medicina ◽

10.3390/medicina43100099 ◽

2007 ◽

Vol 43 (10) ◽

pp. 778

Author(s):

Astra Vitkauskienė ◽

Agnė Giedraitienė ◽

Vytis Dudzevičius ◽

Raimundas Sakalauskas

Keyword(s):

Klebsiella Pneumoniae ◽

Late Onset ◽

Lavage Fluid ◽

Beta Lactamase ◽

Beta Lactamases ◽

Extended Spectrum Beta Lactamase ◽

Extended Spectrum ◽

Extended Spectrum Beta Lactamases ◽

Hospital Acquired Pneumonia ◽

Hospital Acquired

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.

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Seagulls of the Berlengas Natural Reserve of Portugal as Carriers of Fecal Escherichia coli Harboring CTX-M and TEM Extended-Spectrum Beta-Lactamases

Applied and Environmental Microbiology ◽

10.1128/aem.00949-08 ◽

2008 ◽

Vol 74 (23) ◽

pp. 7439-7441 ◽

Cited By ~ 69

Author(s):

Patricia Poeta ◽

Hajer Radhouani ◽

Gilberto Igrejas ◽

Alexandre Gonçalves ◽

Carlos Carvalho ◽

...

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

Antibiotic Resistance Gene ◽

Beta Lactamase ◽

Beta Lactamases ◽

Extended Spectrum Beta Lactamase ◽

Extended Spectrum ◽

Natural Reserve ◽

Class 1 ◽

Extended Spectrum Beta Lactamases

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.

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Characterization of beta-lactamase gene blaPER-2, which encodes an extended-spectrum class A beta-lactamase.

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.40.3.616 ◽

1996 ◽

Vol 40 (3) ◽

pp. 616-620 ◽

Cited By ~ 82

Author(s):

A Bauernfeind ◽

I Stemplinger ◽

R Jungwirth ◽

P Mangold ◽

S Amann ◽

...

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Beta Lactamase ◽

Reading Frame ◽

Beta Lactamases ◽

Class A ◽

Extended Spectrum ◽

The Family ◽

Extended Spectrum Beta Lactamases ◽

Lactamase Gene

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.

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