https://orcid.org/0000-0002-1685-5842
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Abstract
Background
The emergence of carbapenem-resistant and hypervirulent hypermucoviscous Klebsiella pneumoniae strains poses a significant public health challenge. We determined the MDR profiles, antibiotic resistance factors, virulence gene complement, and hypermucoviscous features of 200 clinical K. pneumoniae isolates from two major tertiary care hospitals in Islamabad and Rawalpindi, Pakistan.
Methods
Susceptibility profiling and phenotypic analysis were performed according to the CLSI guidelines. Genetic determinants of antibiotic resistance and virulence were detected by PCR. Biofilm formation analysis was performed by microtiter plate assay.
Results
The isolates displayed a high degree of antibiotic resistance: 36% MDR-CRKP; 38% carbapenem resistance; 55% gentamicin resistance; 53% ciprofloxacin resistance; and 59% aztreonam resistance. In particular, the level of resistance against fosfomycin (22%) and colistin (15%) is consistent with previous reports of increased resistance levels. Combined resistance to carbapenem and colistin was 7%. Genetic factors associated with colistin resistance (mcr-1 and mcr-2 genes) were detected in 12 and 9% of the isolates, respectively. Significant differences in resistance to gentamicin and levofloxacin were observed between the 200 isolates. Many of the isolates harbored genes specifying extended-spectrum and/or carbapenem-resistant β-lactamases: blaCTX-M-15 (46%), blaNDM-1 (39%), and blaOXA-48 (24%). The prevalence of the hypermucoviscous phenotype was 22% and 13% of the MDR isolates carried the rmpA gene (regulator for mucoid phenotype). Key virulence factor genes detected include those encoding: porins (ompK35 and ompK36; at 56 and 55% prevalence, respectively); adhesins (fimH, mrkD, and ycfM; at 19, 18, and 22% prevalence, respectively); and the polysaccharide regulator, bss, at 16% prevalence.
Conclusion
This report highlights carbapenem-resistant K. pneumoniae (CRKP) prevalence, emerging resistance to fosfomycin, and the presence of mcr-1 and mcr-2 in colistin-resistant isolates. Further, the detection of rmpA signifies the prevalence of the hypermucoviscous trait in CRKP clinical isolates from Pakistan.
Acknowledgments
We are thankful to Higher Education Commission of Pakistan for supporting doctoral work of Wajiha Imtiaz.
Abbreviations
MDR, multidrug-resistant; ESBLs, extended-spectrum beta-lactamases; MICs, minimum inhibitory concentration; UTIs, urinary tract infections; ECDC, European Center for Disease Prevention and Control; AmpC, Ampicillinase C; CRE, carbapenem-resistant Enterobacteriaceae; CRKP, carbapenem-resistant Klebsiella pneumoniae; PCR, polymerase chain reaction; HMVKP, hypermucoviscous Klebsiella pneumoniae.
Data Sharing Statement
The data generated from the research work is presented in this article. Any additional information required can be requested from the corresponding author as per ethical guidelines.
Ethics Approval and Consent to Participate
The study was reviewed and granted approval by the Bio-Ethical Committee (BEC) of Quaid-i-Azam University, Islamabad; under the protocol number #BEC-FBS-QAU2019-148. The clinical samples were part of the routine hospital laboratory procedure.
Author Contributions
All authors contributed significantly in conception, study design experimental work, data acquisition analysis and interpretation or in all aspects. Furthermore, all authors took part in drafting, revising and critically reviewing the article and gave final approval of the version being published and agreed on the journal to which article has been submitted and remain accountable for all aspects of the work reported.
Disclosure
The authors declare that they have no conflicts of interest for this work.