AUTHOR=Lascols Christine , Cherney Blake , Conley Andrew B. , Rishishwar Lavanya , Crawford Matthew A. , Morse Stephen A. , Fisher Debra J. , Anderson Kevin , Hodge David R. , Pillai Segaran P. , Hughes Molly A. , Khan Erum , Sue David 

TITLE=Investigation of multidrug-resistant plasmids from carbapenemase-producing Klebsiella pneumoniae clinical isolates from Pakistan

JOURNAL=Frontiers in Microbiology

VOLUME=14

YEAR=2023

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2023.1192097

DOI=10.3389/fmicb.2023.1192097

ISSN=1664-302X

ABSTRACT=<sec><title>Objectives</title><p>The study aim was to investigate multidrug-resistant (MDR) plasmids from a collection of 10 carbapenemase-producing <italic>Klebsiella pneumoniae</italic> clinical isolates identified within the same healthcare institution in Pakistan. Full characterization of the MDR plasmids including structure, typing characteristics, and AMR content as well as determination of their plasmid-based antimicrobial susceptibility profiles were carried out.</p></sec><sec><title>Methods</title><p>Plasmids were isolated from 10 clinical isolates of <italic>Klebsiella pneumoniae</italic>, and from a corresponding set of <italic>Escherichia coli</italic> transconjugants, then sequenced using Nanopore/Illumina technology to generate plasmid hybrid assemblies. Full characterization of MDR plasmids, including determination of next generation sequencing (NGS)-based AMR profiles, plasmid incompatibility groups, and types, was carried out. The structure of MDR plasmids was analyzed using the Galileo AMR platform. For <italic>E. coli</italic> transconjugants, the NGS-based AMR profiles were compared to NGS-predicted AMR phenotypes and conventional broth microdilution (BMD) antimicrobial susceptibility testing (AST) results.</p></sec><sec><title>Results</title><p>All carbapenemase-producing <italic>K. pneumoniae</italic> isolates (carrying either <italic>bla</italic><sub>NDM-1</sub>, or/and <italic>bla</italic><sub>OXA-48</sub>) carried multiple AMR plasmids encoding 34 antimicrobial resistance genes (ARGs) conferring resistance to antimicrobials from 6 different classes. The plasmid incompatibility groups and types identified were: IncC (types 1 and 3), IncFIA (type 26) IncFIB, IncFII (types K1, K2, K7, and K9), IncHI1B, and IncL. None of the <italic>bla</italic><sub>NDM-1</sub> and <italic>bla</italic><sub>ESBL</sub>-plasmids identified in this study were previously described. Most <italic>bla</italic><sub>NDM-1-</sub>plasmids shared identical AMR regions suggesting potential genetic material/plasmid exchange between <italic>K. pneumoniae</italic> isolates of this collection. The majority of NGS-based AMR profiles from the <italic>E. coli</italic> transconjugants correlated well with both NGS-based predicted and conventional AST results.</p></sec><sec><title>Conclusion</title><p>This study highlights the complexity and diversity of the plasmid-based genetic background of carbapenemase-producing clinical isolates from Pakistan. This study emphasizes the need for characterization of MDR plasmids to determine their complete molecular background and monitor AMR through plasmid transmission between multi-resistant bacterial pathogens.</p></sec>