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Characterization of the extended-spectrum b-lactamases and determination of the virulence factors of uropathogenic Escherichia coli strains isolated from children

Charakterisierung der extended spectrum b-Lactamasen (ESBL) und Bestimmung der von Kindern isolierten uropathogenen Virulenzfaktoren von Escherichia coli (E. coli) Stämmen

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Summary

Background and aim

The aim of the study was to characterize ESBL-producing uropathogenic Escherichia coli (UPEC) strains isolated in children. That included the investigation of virulence factors and the analysis of the types of β-lactamases at the molecular genetic level.

Material and methods

During the 2-year study period, 77 ESBL-producing E. coli strains were recovered from urine samples of febrile children with significant bacteriuria hospitalized at one Croatian hospital. Susceptibility of isolates to bactericidal serum activity was tested by Shiller and Hatch method, while adhesin expression was determined by agglutination methods. Characterization of ESBLs was performed by PCR with specific primers for ESBLs and by sequencing of bla ESBL genes. Genotyping of the E. coli isolates was performed by pulsed-field gel electrophoresis (PFGE).

Results

Twenty-seven (35.1 %) and 50 (64.9 %) ESBL-producing UPEC strains were isolated in neonates and infants, respectively. Of 70 strains investigated for the presence of virulence factors, adhesins were detected in 48.6 % strains (8.6 % in the neonate and 40 % in the infants group) giving a statistically significant difference in adhesin expression between the two groups (p < 0.01). Hemolysin was produced by 84.3 %, whereas 70 % of strains were serum-resistant. The bla TEM gene was detected in 22 (28 %) and bla SHV gene in 57 strains (74 %), whereas bla CTX-M gene was detected in only two isolates (2.5%). In ten isolates, bla TEM and bla SHV were simultaneously detected. Sequencing of bla SHV genes revealed that SHV-5 β-lactamase was by far the most prevalent and was found in 51 strains (66 %). The strains were clonally related as demonstrated by PFGE and assigned into ten clusters.

Conclusions

Infection control measures should be employed and the consumption of expanded-spectrum cephalosporins in the hospital should be restricted.

Zusammenfassung

Hintergrund und Ziel der Studie

Ziel der Studie war es, von Kindern isolierte ESBL produzierende uropathogene E. coli (UPEC) Stämme zu charakterisieren und die Typen der β-Lactamasen auf molekular-genetischem Niveau zu analysieren.

Material und Methoden

Während der 2 jährigen Studienperiode wurden 77 ESBL produzierende E coli Stämme aus Harnproben von an einem kroatischen Spital stationär aufgenommenen Kindern isoliert. Mittels der Shiller – Hatch Methode wurde die Empfindlichkeit der Isolate auf Aktivität von bakterizidem Serum getestet. Die Adhesin Expression wurde mit Agglutinationsmethoden untersucht. Die Charakterisierung der ESBL erfolgte mittels PCR mit spezifischen Primers für ESBL und durch Sequenzierung der bla ESBL Gene. Die Gentypisierung der E. coli Isolate wurde mittels puls-feld-gel- Elektrophorese (PGFE) durchgeführt.

Ergebnisse

ESBL produzierende UPEC Kulturen wurden aus Harnproben von 27 (35,1 %) Neugeborenen und von 50 (64,9 %) älteren Kindern isoliert. Von den 70 auf das Vorhandensein von Virulenzfaktoren untersuchten Kulturen wurden bei 48,6 % Adhesine entdeckt (8,6 % bei den Neugeborenen und 40 % bei den Älteren). Die Adhesin Expression beider Gruppen unterschied sich demnach signifikant (p < 0,01). Hämolysin wurde von 84,3 % produziert. Siebzig Prozent der Kulturen waren Serumresistent. Das bla TEM Gen wurde in 22 (28 %) und das bla SHV Gen in 57 (74 %) Kulturen nachgewiesen. Das bla CTX-M Gen konnte aber nur in zwei Isolaten 2,5 % entdeckt werden. Die Sequenzierung der bla SHV Gene zeigte, dass die SHV – 5 β – Lactamase bei weitem am häufigsten war. Sie wurde bei 51 Kulturen (66 %) gefunden. Durch PFGE konnte gezeigt werden, dass die Kulturen Klon-mäßig mit einander verwandt waren.

Schlussfolgerung

Maßnahmen zur Vermeidung einer Infektion sollten ergriffen werden. Der Einsatz von Cephalosporinen mit erweitertem Spektrum sollte im Spitalsbereich eingeschränkt werden.

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Conflict of interest

The authors declare that there is no actual or potential conflict of interest in relation to this article.

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Authors and Affiliations

  1. Department of Microbiology, School of Medicine, University of Zagreb, Salata 3, 10000, Zagreb, Croatia

    Branka Bedenić &  Jasmina Vraneš

  2. Clinical Hospital Center Zagreb, Zagreb, Croatia

    Branka Bedenić

  3. IML red, synlab MVZ Gauting, Asklepios Fachkliniken, Gauting, Germany

    Sabine Hofmann-Thiel Ph.D. & Harald Hoffmann

  4. Department of Microbiology, Split University Hospital, Split, Croatia

    Marija Tonkić &  Anita Novak

  5. Department of Chemistry, Faculty of Natural Sciences, Mathematics and Education, University of Split, Split, Croatia

    Viljemka Bučević-Popovic Ph.D.

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  1. Branka Bedenić
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  2. Jasmina Vraneš
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  3. Sabine Hofmann-Thiel Ph.D.
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Correspondence to Branka Bedenić.

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Bedenić, B., Vraneš, J., Hofmann-Thiel, S. et al. Characterization of the extended-spectrum b-lactamases and determination of the virulence factors of uropathogenic Escherichia coli strains isolated from children. Wien Klin Wochenschr 124, 504–515 (2012). https://doi.org/10.1007/s00508-012-0210-5

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