Next-generation sequencing of plasmid carrying bla_OXA-48 in Klebsiella pneumoniae from Turkey

Düzgün, Azer Özad and Saral, Ayşegül (2019) Next-generation sequencing of plasmid carrying bla_OXA-48 in Klebsiella pneumoniae from Turkey. Acta Microbiologica et Immunologica Hungarica, 66 (2). pp. 261-272. ISSN 1217-8950

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Abstract

A carbapenem-resistant Klebsiella pneumoniae strain was isolated in Turkey in 2012 and bla_NDM-1 and bla_OXA-48 genes were observed in this strain. The aim of this study was to investigate transferability of plasmid bearing bla_OXA-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 bla_OXA-48 within Tn1999.2 transposon without any other antibiotic resistance gene.

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