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Hui Xu,1,2,* Chunxiu Huo,3,* Yao Sun,3,* Yiheng Zhou,4 Yilin Xiong,3 Zinan Zhao,3 Qi Zhou,3 Li Sha,3 Beibei Zhang,5 Yang Chen3 1Department of Clinical Laboratory, The First Affiliated Hospital of Dalian Medical University, Dalian, China; 2Department of Clinical Laboratory, Harbin Medical University Cancer Hospital, Harbin, China; 3Department of Biotechnology, College of Basic Medical Sciences, Dalian Medical University, Dalian, China; 4Institute for Chronic and Non-Communicable Disease Prevention and Control, Dalian Center for Disease Prevention and Control, Dalian, China; 5Laboratory of Pathogenic Biology, Dalian Medical University, Dalian, China *These authors contributed equally to this work Background: Ventilator-associated pneumonia (VAP) is a common nosocomial infection associated with high morbidity due to multidrug-resistant (MDR) pathogens. The purpose of this study was to determine the occurrence of extended-spectrum β-lactamase (ESBL) genes, especially blaCTX-M-15, in Klebsiella pneumoniae (K. pneumoniae)-associated VAP and to investigate the antimicrobial resistance patterns and molecular epidemiological characteristics of K. pneumoniae strains. Materials and methods: From January 2013 to December 2015, we retrospectively collected 89 VAP-causing K. pneumoniae isolates from tertiary-care hospitals in China, among which ESBL-producing strains were assessed for antimicrobial susceptibility. Several antibiotic resistance genes of clinical relevance in K. pneumonia isolates producing ESBL were investigated. Polymerase chain reaction (PCR) and DNA sequencing were employed to characterize the genetic contexts of blaCTX-M-15. Conjugative plasmids carrying blaCTX-M-15 were obtained by mating and further subjected to replicon typing. The genetic relatedness of isolates was assessed by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing. Results: All of the 30 ESBL-producing isolates identified displayed MDR phenotype, with blaSHV, blaCTX-M, blaOXA, and blaTEM detected in 21, 21, 1, and 20 isolates, respectively. blaCTX-M-15 was the most prevalent ESBL gene (19/30, 63.33%), and ISEcp1 was detected 48 bp upstream of 15 blaCTX-M-15 genes. Based on S1-PFGE analyses, 25 isolates exhibited different plasmid profiles, ranging from ~70 to 320 kb. The blaCTX-M-15 with blaTEM and qnr genes and the ISEcp1 element from eight isolates were co-transferrable to recipients via conjugation, with IncFIB, IncFIC, and IncFII being the most prevalent replicons. Twenty different PFGE patterns and 11 sequence types were identified, with ST304 being dominant. Conclusion: This work reports the emergence of blaCTX-M-15 in K. pneumoniae-induced VAP in China. We showed that IncFIB, IncFIC, and/or IncFII plasmids carrying blaCTX-M-15 with blaTEM, qnr resistance genes, and the ISEcp1 element mediate the local prevalence in K. pneumoniae-associated VAP. Keywords: Enterobacteriaceae, CTX-M-15, antibiotic resistance, horizontal gene transfer, conjugation |
