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Plasmid acquired antimicrobial resistance in staphylococcus epidermidis in a peritonitis case
Journal article   Peer reviewed

Plasmid acquired antimicrobial resistance in staphylococcus epidermidis in a peritonitis case

M. Kopczyk, J.P. Ramsay, K. Mulroney, E. Colombi, S. Mowlaboccus, G. Coombs and A. Chakera
Nephrology, Vol.27(S1)
2022
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Abstract

Aim: To sequence the genomes of three longitudinal S. epidermidis isolates (C099, C100 and C101) obtained from a patient with peritonitis. Background: Coagulase-negative staphylococci such as S. epidermidis are common colonizers on human skin, however as opportunistic pathogens they can cause serious infections. S. epidermidis is resistant to a range of antibiotics such as methicillin and tetracycline and most antimicrobial resistance (AMR) genes are plasmid encoded. Plasmids can facilitate the rapid spread of AMR between bacteria and we have observed this in S. epidermidis isolates obtained from a treatment resistant peritonitis case. Method: C099 was isolated from the patient on day 1 of infection, C100 was obtained on day 6 and C101 was obtained on day 15. DNA was extracted using lysostaphin lysis of cells followed by the use of the Monarch® Genomic DNA purification kit. Extracted DNA was checked for quality and sequenced using the Illumina NextSeq and Oxford Nanopore MinION. Replicons were confirmed using gel electrophoresis. Genomes were assembled using long nanopore sequence reads using Flye. Final assemblies were polished with both long reads using Racon and with short reads (Illumina) using Pilon. The broth micro-dilution (BMD) method was performed using Sensititre™ Custom plates according to manufacturer's instructions (Thermo Scientific™). Results: A total of five plasmids were identified in C099 and six in C100 and C101. Sequencing confirmed the additional plasmid in C100 and C101 contained the ermC gene. Both C100 and C101 were resistant to erythromycin using the BMD method. Conclusions: The rapid acquisition of erythromycin resistance highlights the issue of increasing AMR in bacteria. Sequencing can be used for surveillance and to further study the emergence of AMR in peritonitis cases.

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