Klebsiella pneumoniae multidrug-resistant (MDR) infections represent a global health concern and present significant clinical hurdles in both community as well as hospital settings, the development of resistant in Gram-negative strains (G -) occurs more rapidly in comparison with Gram-positive strains (G+). The present study aims to genetically investigate Beta-lactamase enzymes coding genes in isolates of K. pneumoniae MDR producing Extended spectrum β- lactamase enzymes (ESBLs), Metallo β- lactamase enzymes (MBLs) and (AmpC) using whole genome sequencing (WGS Isolates) were identified, and antibiotic sensitivity test for ten antibiotics was conducted using VITEK2.Whole Genome Sequencing (WGS) was conducted on two isolates resistant to Cephalosporins and Carbapenems utilizing the Illumina MiSeq technology. The sequences were compared to the antimicrobial resistance (AMR) gene database following the annotation of the assembled draft genomes. Results: Many bacteria show several resistance genes have been identified that facilitate the manufacture of β-lactamases through WGS, which include (ESBLs, MBLs, and AmpC) genes, as (blaSHV-28/79 , blaTEM-1B, blaCTX-M-15, blaCMY-6, blaNDM-1/2, blaMOX-1, blaDHA-1). Furthermore, Quinolone resistance includes (OqxA, OqxB, qnrS1, qnrB4, parC, acrRMoreover), genes that encoding aminoglycosides modification enzymes (rpsL, aac(3')-lla, aac(6')-Ib3, aadA5, rmtC, aph(3')-la), trimethoprim- sulfamethoxazole (dfrA17, sul1), Tetracycline (tetA, tetD, ramR), Fosfomycin (fosA6), chloramphenicol (catA2) resistance genes, while other genes that endow Macrolide resistance (mphA), and carbapenem resistance genes (OmpK35, OmpK36,OmpK37). The molecular characterization of isolates can be reconstructed using WGS thorough analysis.
Z. H. Mahmud et al., “Healthcare Facilities as Potential Reservoirs of Antimicrobial Resistant Klebsiella pneumoniae: An Emerging Concern to Public Health in Bangladesh,” Pharmaceuticals, vol. 15, no. 9, p. 1116, 2022.
U. Kilic, M. Koroglu, M. Olmez, and M. Altindis, “Investigation of the In Vitro Effectiveness of Aztreonam/Avibactam, Colistin/Apramycin, and Meropenem/Apramycin Combinations Against Carbapenemase-Producing, Extensively Drug-Resistant Klebsiella pneumoniae Strains,” Microbial Drug Resistance, vol. 26, no. 11, pp. 1291-1297, 2020.
J. Noster, P. Thelen, and A. Hamprecht, “Detection of Multidrug-Resistant Enterobacterales - From ESBLs to Carbapenemases,” Antibiotics, vol. 10, no. 9, p. 1140, 2021.
S. Alfei and A. M. Schito, “Positively Charged Polymers as Promising Devices Against Multidrug Resistant Gram-Negative Bacteria: A Review,” Polymers, vol. 12, no. 5, p. 1195, 2020.
H. Karadottir et al., “Klebsiella pneumoniae Expressing VIM-1 Metallo-β-Lactamase Is Resensitized to Cefotaxime via Thiol-Mediated Zinc Chelation,” Infection and Immunity, vol. 88, no. 1, pp. 10.1128/IAI.00756-19, 2019.
A. Y. Shareef and S. L. Saleem, “Detection of AmpC Enzyme in Gram Negative Bacteria by Phenotypic and Molecular Methods,” 2021.
Q. Zhou, M. Tang, X. Zhang, J. Lu, X. Tang, and Y. Gao, “Detection of AmpC β-Lactamases in Gram-Negative Bacteria,” Heliyon, vol. 8, no. 12, 2022.
A. V. Bassenden et al., “Structural and Phylogenetic Analyses of Resistance to Next-Generation Aminoglycosides Conferred by AAC (2′) Enzymes,” Scientific Reports, vol. 11, no. 1, p. 11614, 2021.
P. Wayne, “Clinical and Laboratory Standards Institute. Performance Standards for Antimicrobial Susceptibility Testing,” 2011.
V. Mishra, V. L. Nag, R. Tandon, and S. Awasthi, “Response Surface Methodology-Based Optimisation of Agarose Gel Electrophoresis for Screening and Electropherotyping of Rotavirus,” Applied Biochemistry and Biotechnology, vol. 160, no. 8, pp. 2322-2331, 2010.
“BBMap & BBTools: The Official Suite of Bioinformatics Tools,” [Online]. Available: https://jgi.doe.gov/data-and-tools/software-tools/bbtools/bb-tools-user-guide/bbduk-guide/.
A. Bankevich et al., “SPAdes: A New Genome Assembly Algorithm and Its Applications to Single-Cell Sequencing,” Journal of Computational Biology, vol. 19, no. 5, pp. 455-477, 2012.
C. Camacho et al., “BLAST+: Architecture and Applications,” BMC Bioinformatics, vol. 10, no. 1, p. 421, 2009.
E. Ugbo et al., “Prevalence of blaTEM, blaSHV, and blaCTX-M Genes Among Extended Spectrum Beta-Lactamase-Producing Escherichia coli and Klebsiella pneumoniae of Clinical Origin,” Gene Reports, vol. 21, p. 100909, 2020.
S. G. Rimoldi et al., “Whole Genome Sequencing for the Molecular Characterization of Carbapenem-Resistant Klebsiella pneumoniae Strains Isolated at the Italian ASST Fatebenefratelli Sacco Hospital, 2012-2014,” BMC Infectious Diseases, vol. 17, no. 1, p. 666, 2017.
K. Lai, Y. Ma, L. Guo, J. An, L. Ye, and J. Yang, “Molecular Characterization of Clinical IMP-Producing Klebsiella pneumoniae Isolates from a Chinese Tertiary Hospital,” Annals of Clinical Microbiology and Antimicrobials, vol. 16, no. 1, p. 42, 2017.
S. Li et al., “First Report of blaCTX-M-167, blaSHV-1, and blaTEM-1B Carrying Klebsiella pneumoniae Showing High-Level Resistance to Carbapenems,” Frontiers in Microbiology, vol. 13, p. 916304, 2022.
M. Biggel, K. Zurfluh, A. Treier, M. Nüesch-Inderbinen, and R. Stephan, “Characteristics of fosA-Carrying Plasmids in E. coli and Klebsiella spp. Isolates Originating from Food and Environmental Samples,” Journal of Antimicrobial Chemotherapy, vol. 76, no. 8, pp. 2004-2011, 2021.
X. Zhang et al., “High-Level Aminoglycoside Resistance in Human Clinical Klebsiella pneumoniae Complex Isolates and Characteristics of armA-Carrying IncHI5 Plasmids,” Frontiers in Microbiology, vol. 12, p. 636396, 2021.
Q. Xu et al., “RamA Upregulates Multidrug Resistance Efflux Pumps AcrAB and OqxAB in Klebsiella pneumoniae,” International Journal of Antimicrobial Agents, vol. 57, no. 2, p. 106251, 2021.
S.-K. Chiu et al., “Roles of ramR and tet(A) Mutations in Conferring Tigecycline Resistance in Carbapenem-Resistant Klebsiella pneumoniae Clinical Isolates,” Antimicrobial Agents and Chemotherapy, vol. 61, no. 8, pp. 10.1128/AAC.00391-17, 2017.
A. Rocker et al., “Global Trends in Proteome Remodeling of the Outer Membrane Modulate Antimicrobial Permeability in Klebsiella pneumoniae,” mBio, vol. 11, no. 2, pp. 10.1128/mBio.00603-20, 2020.
O. H. Moghnia and N. A. Al-Sweih, “Whole Genome Sequence Analysis of Multidrug Resistant Escherichia coli and Klebsiella pneumoniae Strains in Kuwait,” Microorganisms, vol. 10, no. 3, p. 507, 2022.
S. S. Mohamed, A. M. A. Dayem, M. L. Sakr, and I. A. Dwedar, “The Effect of Administration of Fosfomycin in the Management of Ventilator-Associated Pneumonia,” The Egyptian Journal of Chest Diseases and Tuberculosis, vol. 67, no. 3, pp. 318-322, 2018.
