AUTHOR=Zeng Lingyi , Zhang Jisheng , Hu Kewang , Li Jie , Wang Jianmin , Yang Chengru , Huang Wan , Yin Lining , Zhang Xiaoli 

TITLE=Microbial Characteristics and Genomic Analysis of an ST11 Carbapenem-Resistant Klebsiella pneumoniae Strain Carrying blaKPC−2 Conjugative Drug-Resistant Plasmid

JOURNAL=Frontiers in Public Health

VOLUME=9

YEAR=2022

URL=https://www.frontiersin.org/journals/public-health/articles/10.3389/fpubh.2021.809753

DOI=10.3389/fpubh.2021.809753

ISSN=2296-2565

ABSTRACT=<sec><title>Background</title><p>The sequence type 11 (ST11) carbapenem-resistant <italic>Klebsiella pneumoniae</italic> (CRKP) carrying <italic>bla</italic><sub><italic>KPC</italic>−2</sub> has been widespread all over the world, and it has been reported frequently in China. The <italic>bla</italic><sub><italic>KPC</italic>−2</sub> located on the mobile genetic element brings tremendous pressure to control the spread and outbreak of resistant bacteria. Whole-genome sequencing (WGS) technology can comprehensively and in-depth display the molecular characteristics of drug-resistant bacteria, providing a basis for evaluating the genetic diversity within the CRKP genome.</p></sec><sec><title>Methods</title><p>The ST11 CRKP in this study was collected in the intensive care unit of a major teaching hospital. PCR and Sanger sequencing confirmed the existence of <italic>bla</italic><sub><italic>KPC</italic>−2</sub>. The AST-GN card and the microbroth dilution test were used for antimicrobial susceptibility testing. The transferability of plasmid was verified by a conjugation test. The whole genome is sequenced using the Illumina HiSeq short-read and Oxford Nanopore long-read sequencing technology.</p></sec><sec><title>Results</title><p>The studied strain was named CRKP63, which is a multi-drug resistance bacteria, which carries <italic>bla</italic><sub><italic>KPC</italic>−2</sub> and <italic>bla</italic><sub><italic>SHV</italic>−182</sub>. Its genome consists of a circular chromosome of 5,374,207 bp and an IncFII plasmid named pKPC-063001 of 359,625 bp. In the drug-resistant plasmid pKPC-063001, the key carbapenem resistance gene <italic>bla</italic><sub><italic>KPC</italic>−2</sub> was located in the genetic context with insertion sequence IS<italic>Kpn27</italic> upstream and IS<italic>Kpn6</italic> downstream and bracketed by IS<italic>26</italic>. The three copies of the IS<italic>26</italic>–IS<italic>Kpn27</italic>–<italic>bla</italic><sub><italic>KPC</italic>−2</sub>–IS<italic>Kpn6</italic>–IS<italic>26</italic> unit were present in tandem. <italic>bla</italic><sub><italic>KPC</italic>−2</sub> can be transferred horizontally between other species by conjugation, the complete type IV secretion system (T4SS) structure helps to improve the adaptability of bacteria to the external environment, strengthen the existence of drug-resistant bacteria, and accelerate the spread of drug resistance.</p></sec><sec><title>Conclusion</title><p>High-throughput sequencing has discovered the different surrounding environments of <italic>bla</italic><sub><italic>KPC</italic>−2</sub>, which provides a new idea for further revealing the transmission and inheritance of <italic>bla</italic><sub><italic>KPC</italic>−2</sub> at the molecular level. In order to control the further spread and prevalence of drug-resistant bacteria, we should pay close attention to the changes in the genetic environment of <italic>bla</italic><sub><italic>KPC</italic>−2</sub> and further study the transcription and expression of T4SS.</p></sec>