Klebsiella pneumoniae ST307 with OXA-181: threat of a high-risk clone and promiscuous plasmid in a resource-constrained healthcare setting.

INTRODUCTION: Klebsiella pneumoniae with OXA-48-like enzymes were introduced into Tshwane Tertiary Hospital (TTH) (Pretoria, South Africa) during September 2015, causing nosocomial outbreaks. METHODS: PCR methodologies and WGS were used to characterize K. pneumoniae with carbapenemases (n = 124) from TTH (July 2015-December 2016). RESULTS: PCR was used to track K. pneumoniae ST307 with OXA-181 among 60% of carbapenemase-positive isolates in different wards/units over time and showed the transmission of IncX3 plasmids to other K. pneumoniae clones. WGS identified different ST307 clades: 307_OXA181 (consisting of two lineages, A and B) with OXA-181 on IncX3 plasmids (named p72_X3_OXA181) and clade 307_VIM with VIM-1 on IncFII plasmids. Clade 307_OXA181 lineage B was responsible for the rapid increase and transmission of OXA-181 K. pneumoniae in various wards/units throughout TTH, while the numbers of clade 307_OXA181 lineage A and clade 307_VIM remained low. Separate outbreaks were due to K. pneumoniae ST17 and ST29 with p72_X3_OXA181 plasmids. CONCLUSIONS: The high-risk clone K. pneumoniae ST307 with OXA-181 rapidly spread to different wards/units despite infection and prevention measures. ST307 clades and lineages seemingly acted differently in outbreak situations. This study also highlighted the threat of promiscuous plasmids such as p72_X3_OXA181.

Evaluation of an In-House Colistin NP Test for Use in Resource-Limited Settings.

Colistin has become increasingly important in the treatment of multidrug-resistant Gram-negative bacteria. Resistance to colistin has emerged globally, necessitating the need for an accurate method to detect colistin resistance. The colistin NP test has shown promise as a rapid screening assay for colistin resistance. This study compared the performance of an in-house-prepared colistin NP test against broth microdilution (BMD) as the gold standard and against Etest (bioMérieux, Marcy l'Etoile, France) as an alternative method. A total of 215 stored Enterobacteriaceae isolates were evaluated, of which 159 were resistant and 56 susceptible to colistin by BMD. The categorical agreement of the colistin NP test with BMD was found to be 98.1%, compared to 87.9% for the Etest. One major error was detected with both the colistin NP test and the Etest. Three very major errors were detected with the colistin NP test compared to 25 with the Etest. This resulted in a major error rate of 1.8% for both the colistin NP test and the Etest and a very major error rate of 1.9% and 15.7% for the colistin NP test and the Etest, respectively. The colistin NP test compared satisfactorily to the BMD reference method in determining colistin susceptibility. The colistin NP test is a rapid, inexpensive screening method for colistin resistance, especially in resource-limited settings.