Emergence and nosocomial spread of carbapenem-resistant OXA-232-producing Klebsiella pneumoniae in Brunei Darussalam.

Abstract

Carbapenem-resistant Enterobacteriaceae (CRE) are identified as a major global health concern. The success of CRE is facilitated by the emergence, acquisition and spread of successful clones carrying plasmid-encoded resistance genes. In this study, an outbreak of carbapenem-resistant Klebsiella pneumoniae (CRKP) infections in patients hospitalised in Brunei Darussalam was investigated. Over a 3-month period (May-July 2015), five multidrug-resistant K. pneumoniae were recovered from individual patients admitted to intensive care units at two hospitals (RIPAS and PMMPMHAB) in Brunei. Antimicrobial susceptibility was determined by broth microtitre dilution using a Micronaut-S β-lactamase VII kit or by Etest. Carbapenemase production was confirmed using the RAPID CARB Blue screen, and classes A-D β-lactamases were detected by multiplex PCR. Molecular typing was performed by random amplified polymorphic DNA (RAPD) PCR and multilocus sequence typing (MLST), with associated virulence and capsular types identified by PCR and sequencing. Plasmids were extracted, sized and characterised by PCR-based replicon typing. All isolates were resistant to cephalosporins, carbapenems, aminoglycosides, quinolones and sulfonamides but remained susceptible to polymyxins. Isolates were indistinguishable by RAPD-PCR and all belonged to sequence type (ST231). Resistance was due to the production of OXA-232 and CTX-M-15 β-lactamases, with the blaOXA-232 carbapenemase gene located on a ColE-like plasmid. This is the first report of plasmid-encoded OXA-232-producing CRKP in Brunei hospitals. All isolates were members of ST231, which may be representatives of a high-risk CRKP clone disseminating across Southeast Asia.