PCR Detection of Oxacillinases in Bacteria.
Oxacillinases (OXA) have been mostly described in Enterobacteriaceae, Acinetobacter, and Pseudomonas species. Recent years have witnessed an increased prevalence of intrinsic and/or acquired β-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.
['Mlynarcik|Patrik|P|', 'Chalachanova|Andrea|A|', 'Vagnerovă|Iva|I|', 'Holy|Ondrej|O|', 'Zatloukalova|Simona|S|', 'Kolar|Milan|M|']
D000818:Animals D000900:Anti-Bacterial Agents D001426:Bacterial Proteins D002645:Chickens D018153:Czech Republic D017931:DNA Primers D016972:Enterobacter cloacae D004926:Escherichia coli D015870:Gene Expression D016905:Gram-Negative Bacterial Infections D006801:Humans D007711:Klebsiella pneumoniae D008460:Meat D008826:Microbial Sensitivity Tests D060885:Multiplex Polymerase Chain Reaction D010406:Penicillins D010802:Phylogeny D001618:beta-Lactamases
PCR oxacillinases primers antibiotic resistance