Emergence and molecular characterization of multidrug-resistant emKlebsiella pneumoniae/em isolates harboring embla/emsubCTX-M-15/sub extended-spectrum β-lactamases causing ventilator-associated pneumonia in China

摘要

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 bla CTX-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 bla CTX-M-15. Conjugative plasmids carrying bla CTX-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 bla SHV, bla CTX-M, bla OXA, and bla TEM detected in 21, 21, 1, and 20 isolates, respectively. bla CTX-M-15?was the most prevalent ESBL gene (19/30, 63.33%), and IS Ecp1 was detected 48 bp upstream of 15 bla CTX-M-15?genes. Based on S1-PFGE analyses, 25 isolates exhibited different plasmid profiles, ranging from ~70 to 320 kb. The bla CTX-M-15?with blaTEM and qnr genes and the IS Ecp1 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 bla CTX-M-15?in K. pneumoniae -induced VAP in China. We showed that IncFIB, IncFIC, and/or IncFII plasmids carrying bla CTX-M-15?with bla TEM, qnr resistance genes, and the IS Ecp1 element mediate the local prevalence in K. pneumoniae -associated VAP.