Sürdürülebilir Kalkınma Amaçları

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• MakaleAçık Erişim2021/r/gumushane/14928
  Antibiotic induced biofilm formation of novel multidrug resistant Acinetobacter baumannii ST2121 clone

  Çimen, Müberra; Düzgün, Azer Özad

  The aim of this study was to identify antimicrobial resistance and virulence factor genes exhibited by multidrug resistant (MDR) Acinetobacter baumannii, to analyze biofilm formation and to investigate clonal subtypes of isolate. Whole genome sequencing was done by Illumina NovaSeq 6,000 platform and multilocus sequence typing (MLST) was performed by Oxford and Pasteur typing schemes. Influence of imipenem and levofloxacin on biofilm formation was investigated in 96-well plates at 3 replicates. The strain was found to carry OXA-23, OXA-51-like, AmpC and TEM-1 beta-lactamases. The sequence of the blaOXA-51-like gene has been identified as a blaOXA-66. According to Pasteur MLST scheme the strain displayed ST2 allelic profile. However, based on Oxford MLST scheme this strain represents the new ST2121, as the gdhB gene has a single allelic mutation namely, the gdhB-227. It was determined that MDR isolate carried bap, basABCDFGHIJ, csuA/BABCDE, bauABCDEF, plcD, pgaABCD, entE, barAB, ompA, abaIR, piT2EAFTE/AUBl, fimADT, cvaC, bfmR, bfmS virulence genes. In our study imipenem induced the highest biofilm formation at a concentration of 32 mg/ml and levofloxacin at a concentration of 16 mg/ml. In conclusion, we detected a new MDR A. baumannii ST2121 clone harboring blaOXA-66 gene that has been reported for the first time in Turkey.

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  İncele
• MakaleAçık Erişim2018/r/gumushane/15074
  Next-generation sequencing of plasmid carrying blaOXA-48 in Klebsiella pneumoniae from Turkey

  Düzgün, Azer Özad; Saral, Ayşegül

  A carbapenem-resistant Klebsiella pneumoniae strain was isolated in Turkey in 2012 and blaNDM-1 and blaOXA-48 genes were observed in this strain. The aim of this study was to investigate transferability of plasmid bearing blaOXA-48 in K. pneumoniae and to use whole-genome sequencing in order to understand the genetic context of plasmid. K. pneumoniae strain was used as donor in conjugation experiments. Antibiotic susceptibility profile of selected transconjugant was determined. Plasmid was isolated from transconjugant colony and was named as pKPT. Complete sequencing of the pKPT was conducted using a next-generation sequencing. Annotation of the contigs was performed using the Geneious R9, followed by finding open reading frames (ORFs) with selected web-based tools. BLAST analysis was performed at the NCBI BLAST server to determine genes showing more than 90% similarity with these ORFs. Results of antibiotic susceptibility test showed that transconjugant colony was resistant to ampicillin/ sulbactam, piperacillin, and piperacillin/tazobactam. The pKPT plasmid had a length of 45,217 bp and an average G + C content of 49%. Blast analysis revealed that pKPT was included in the IncL/M incompatibility group. The pKPT was found to contain blaOXA-48 within Tn1999.2 transposon without any other antibiotic resistance gene.

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  İncele
• MakaleAçık Erişim2018/r/gumushane/11834
  Effect of amino acid substitution in New Delhi metallo-β-lactamase on carbapenem susceptibility

  Düzgün, Azer Özad

  The aim of this work was investigation of clinically important amino acid substitutions of NDM-1 variants. A blaNDM-1 gene was cloned into expression vector pET100/D-TOPO. The sequence of NDM-1 variants with substituted amino acids was determined by ClustalW program. A pET100/D-TOPO + blaNDM-1 was used to generate the alanine mutations at different positions, such as NDM-2 (P28A), NDM-3 (D95A), NDM-4 (M154A), NDM-5 (V88A), NDM-7 (D130A), and NDM-9 (E152A). The mutant variants were transformed into Escherichia coli DH5α. Changes in the activities of alanine mutation variants were determined by E-test. All samples had 32 μg/ml MIC values against ampicillin. The 28th amino acid mutation sample had the highest MIC value against ceftazidime, whereas decreased MIC value for piperacillin. It was observed that the resistance to imipenem was increased in mutant variants D95A, M154A, D130A, and E152A, comparing with P28A and V88A. It was found that NDM-1 has 0.64 μg/ml and the 130th amino acid mutation sample has 0.75 μg/ml meropenem MIC value.

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