Identification of IS1R and IS10R elements inserted into ompk36 porin gene of two multidrug-resistant Klebsiella pneumoniae hospital strains

Abstract

Hospital Klebsiella pneumoniae strains (n = 196) were collected in 2012-16 from the patients of a Moscow neurosurgical intensive care unit. Klebsiella pneumoniae strains were multidrug-resistant and carried beta-lactamase genes blaSHV (97.4% of strains), blaCTX-M (84.7%), blaTEM (56.1%), blaOXA-48-like (49.0%) and blaNDM-1 (one strain), class 1 integrons (43.4% of strains) and porin protein ompK36 gene (100% of strains). The ompK36 porin protein gene disruption by insertion sequence (IS) elements and OmpK36 production loss in two strains were detected in this study. Outer membrane proteins were isolated according to Carlone et al. (Rapid microprocedure for isolating detergent-insoluble outer membrane proteins from Haemophilus species. J Clin Microbiol 1986;24:330-2). The IS10R element belonging to the IS4 family, IS10 group was detected at the position of the 41st nucleotide of the ompK36 gene in K. pneumoniae strain KPB-2304K/15 (the first report for a certain IS element in K. pneumoniae). The IS1R element belonging to the IS1 family was identified at the position of the 86th nucleotide of the ompK36 gene in the K. pneumoniae strain KPB-367K/15 (novel insertion site for IS1 element into ompK36 gene). DNA transfer of the intact ompK36 gene into the strain KPB-367K/15 by vector plasmid restored OmpK36 porin protein production and resulted in a decrease of imipenem minimal inhibitory concentration. Such data confirm the importance of IS elements in ongoing multidrug-resistant evolution in hospital Klebsiella.

Targeting Plasmids to Limit Acquisition and Transmission of Antimicrobial Resistance

“…In colistin-resistant strains of Klebsiella sp., alteration of mgrB and phoP gene sequences by different IS, such as ISKpn14, ISKpn28, IS903, IS5, and IS3, can sometimes induce a pandrug-resistance phenotype (Giordano et al, 2018;Uz Zaman et al, 2018). IS also play a vital role in carbapenem resistance through a mechanism similar to that of colistin resistance but involving the inactivation of oprD and omp genes (Lev et al, 2017;Bocharova et al, 2019). The oprD gene is inactivated by the insertion of ISPpu-21 (Shariati et al, 2018).…”

Comparative analysis of Klebsiella pneumoniae strains isolated in 2012–2016 that differ by antibiotic resistance genes and virulence genes profiles

“…Resistance to beta-lactams detected for the strains that have no beta-lactamase genes indicate the presence of additional molecular mechanisms, possibly other types of beta-lactamases, porin mutations and/or efflux pumps (Table 1). For example, recently we described K. pneumoniae strains carrying the insertions of IS1R and IS10R elements that inactivated the ompK36 gene, resulting in loss of the OmpK36 porin and raising resistance to imipenem [29]. Class 1 integrons detected in 38% of K. pneumoniae strains undoubtedly contributed to the resistance phenotype of these strains.…”

The Impact of Insertion Sequences on O-Serotype Phenotype and Its O-Locus-Based Prediction in Klebsiella pneumoniae O2 and O1

“…The hyperepidemic clone ST258, characterized by the notorious production of KPC-type carbapenemases and extensive drug resistance, has more ISs than an average K. pneumoniae isolate of another ST [ 30 , 31 ]. Several IS types are especially frequent in ST258, such as IS Kpn26 [ 30 , 32 ]. In our study, the majority of O2 isolates identified belonged to ST258 (≈70%), and IS Kpn26 was commonly found in these (≈50%).…”