Exploring the phenotype and genotype of multi-drug resistant Klebsiella pneumoniae harbouring bla(CTX-M) group extended-spectrum β-lactamases recovered from paediatric clinical cases in Shenzhen, China

BACKGROUND: Emergence and spread of β-lactamase resistant Klebsiella pneumoniae have posed a serious threat, especially in paediatric patients globally. The present study focuses on explore drug resistance profile and molecular characterization of carbapenemase and extended-spectrum β-lactamase producing K. pneumoniae isolated from paediatric patients in Shenzhen, China. METHODS: Present study, a total of 31 isolates of multi-drug resistant K. pneumoniae were collected from Shenzhen Children’s Hospital, China during Jan 2014 to December 2015. ESBLs production was confirmed by using the combination disc diffusion method followed by antimicrobial susceptibility. In addition, β-lactamase encoding genes were determined by PCR assay and sequencing. The genotypic diversity and phylogenetic relationship were determined by multi-locus sequence typing (MLST) method and pulsed-field gel electrophoresis (PFGE). RESULTS: We examined 31, unique K. pneumoniae isolates collected from 2014 and 2015 in Shenzhen Children’s Hospital, China. All the 31 isolates 100% were resistant to ceftazidime, ertapenem, ampicillin, cefazolin and ampicillin-sulbactam followed by ceftriaxone 94% (n = 29), aztreonam 89% (n = 26), cefepime 84% (n = 26), nitrofurantoin 75% (n = 24), piperacillin 52% (n = 16), and levofloxacin 49% (n = 15). Of the 31 β-lactamase gene coding isolates, bla(CTX-M) was mainly detected in about 100% (n = 31), followed by bla(KPC) 71% (n = 22), bla(SHV) 61% (n = 19), bla(NDM) 25% (n = 8), bla(CYM) 13% (n = 4), bla(OXA-48) 9% (n = 3), bla(GES) 9% (n = 3) and bla(TEM) 6% (n = 2). Seventeen distinct sequences type were observed with ST20 being mostly identified 16% (n = 5). Pulsed-field gel electrophoresis (PFGE) typing showed that identical profile for the isolates recovered from the Department of Intensive Care Unit and Department of Neurology of our hospital. Plasmid replicon typing result indicates the presence of IncFIS type as highest in all isolates as 61% (n = 19), followed by IncFIB 23% (n = 7), IncFIA and IncFIC 16% (n = 5) each. CONCLUSION: Our study reports the occurrence and spread of extended β-lactamase K. pneumoniae ST20 and ST2407 for the first time, in Shenzhen, particularly in paediatric patients. To prevent and control the infection by limiting the spread of infection-causing organisms it is very crucial to detect the presence of resistant genes at an early stage.