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Microb Drug Resist ; 26(9): 1023-1037, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32212994

RESUMO

Oxacillinases (OXA) have been mostly described in Enterobacteriaceae, Acinetobacter, and Pseudomonas species. Recent years have witnessed an increased prevalence of intrinsic and/or acquired ß-lactamase-producing Acinetobacter in food-producing animals. This study was conducted to assess the prevalence of OXA among selected bacterial species and to characterize these enzymes by in silico analysis. Screening of OXA was performed by PCR amplification using specific pairs of oligonucleotides. Overall, 40 pairs of primers were designed, of which 6 were experimentally tested in vitro. Among 49 bacterial isolates examined, the presence of blaOXA-1-like genes was confirmed in 20 cases (41%; 19 times in Klebsiella pneumoniae and once in Enterobacter cloacae). No OXA were found in animal isolates. The study results confirmed the specificity of the designed oligonucleotide pairs. Furthermore, the designed primers were found to possess the ability to specifically detect 90.2% of all OXA. These facts suggest that the in silico and in vitro tested primers could be used for single or multiplex PCR to screen for the presence of OXA in various bacteria, as well as to monitor their spread. At the same time, the presence of conserved characteristic amino acids and motifs was confirmed by in silico analysis of sequences of representative members of OXA.


Assuntos
Proteínas de Bactérias/genética , Enterobacter cloacae/genética , Escherichia coli/genética , Infecções por Bactérias Gram-Negativas/veterinária , Klebsiella pneumoniae/genética , Reação em Cadeia da Polimerase Multiplex/métodos , beta-Lactamases/genética , Animais , Antibacterianos/farmacologia , Proteínas de Bactérias/metabolismo , Galinhas/microbiologia , República Tcheca/epidemiologia , Primers do DNA/síntese química , Primers do DNA/metabolismo , Enterobacter cloacae/classificação , Enterobacter cloacae/efeitos dos fármacos , Enterobacter cloacae/enzimologia , Escherichia coli/classificação , Escherichia coli/efeitos dos fármacos , Escherichia coli/enzimologia , Expressão Gênica , Infecções por Bactérias Gram-Negativas/diagnóstico , Infecções por Bactérias Gram-Negativas/epidemiologia , Infecções por Bactérias Gram-Negativas/microbiologia , Humanos , Klebsiella pneumoniae/classificação , Klebsiella pneumoniae/efeitos dos fármacos , Klebsiella pneumoniae/enzimologia , Carne/microbiologia , Testes de Sensibilidade Microbiana , Penicilinas/farmacologia , Filogenia , beta-Lactamases/metabolismo
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