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Microb Drug Resist ; 26(9): 1023-1037, 2020 Sep.
Article in English | MEDLINE | ID: mdl-32212994

ABSTRACT

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.


Subject(s)
Bacterial Proteins/genetics , Enterobacter cloacae/genetics , Escherichia coli/genetics , Gram-Negative Bacterial Infections/veterinary , Klebsiella pneumoniae/genetics , Multiplex Polymerase Chain Reaction/methods , beta-Lactamases/genetics , Animals , Anti-Bacterial Agents/pharmacology , Bacterial Proteins/metabolism , Chickens/microbiology , Czech Republic/epidemiology , DNA Primers/chemical synthesis , DNA Primers/metabolism , Enterobacter cloacae/classification , Enterobacter cloacae/drug effects , Enterobacter cloacae/enzymology , Escherichia coli/classification , Escherichia coli/drug effects , Escherichia coli/enzymology , Gene Expression , Gram-Negative Bacterial Infections/diagnosis , Gram-Negative Bacterial Infections/epidemiology , Gram-Negative Bacterial Infections/microbiology , Humans , Klebsiella pneumoniae/classification , Klebsiella pneumoniae/drug effects , Klebsiella pneumoniae/enzymology , Meat/microbiology , Microbial Sensitivity Tests , Penicillins/pharmacology , Phylogeny , beta-Lactamases/metabolism
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