| Summary: | The spread of carbapenem-resistant <i>Escherichia coli</i> and <i>Klebsiella pneumoniae</i> is a global concern. The management of infections caused by multidrug resistance (MDR) isolates poses substantial clinical challenges in both hospitals and communities. This study aimed to investigate the genetic characteristics and variations of MDR <i>E. coli</i> and <i>K. pneumoniae</i> isolates. Bacterial identification and antibiotic susceptibility testing against 19 antibiotics were performed by standard methods. Whole genome sequencing (WGS) was carried out on eight carbapenem-resistant isolates using an Illumina MiSeq platform. The assembled draft genomes were annotated, then sequences were blasted against antimicrobial resistance (AMR) genes database. WGS detected several resistance genes mediating the production of β-lactamases, including carbapenems and extended-spectrum β-lactamase genes as <i>(bla<sub>OXA-1/-48</sub></i>, <i>bla</i><sub>KPC-2/-29</sub>, <i>bla</i><sub>CMY-4/-6</sub>, <i>bla</i><sub>SHV-11/-12</sub>, <i>bla</i><sub>TEM-1</sub>, <i>bla</i><sub>CTX-M-15</sub>, <i>bla</i><sub>OKP-B</sub>, <i>bla</i><sub>ACT</sub> and <i>bla</i><sub>EC</sub>). Furthermore quinolone resistance including <i>oqxA/oqxB</i>, <i>aac(6′)-Ib-cr5</i>, <i>gyrA_D87N</i>, <i>gyrA_S83F</i>, <i>gyrA_S83L</i>, <i>parC_S80I</i>, <i>parE_S458A</i>, <i>parE_I355T</i>, <i>parC_S80I</i>, and <i>qnrB1</i>. In addition to aminoglycoside modifying enzymes genes (<i>aph(6)-Id</i>, <i>aph(3</i>″<i>)-Ib</i>, <i>aac(3)-IIa</i>, <i>aac(6′)-Ib</i>, <i>aadA1</i>, <i>aadA2</i> and <i>aadA5</i>), trimethoprim-sulfamethoxazole (<i>dfrA12/A14/A17 and sul1/sul2</i>), tetracycline (<i>tetA</i> and <i>tetB</i>), fosfomycin (<i>fosA and uhpT_E350Q</i>) resistance genes, while other genes were detected conferring chloramphenicol (<i>floR</i>, <i>catA2</i>, and efflux pump <i>cmIA5</i>), macrolides resistance (<i>mph(A) and erm(B)</i>, and quaternary ammonium efflux pump <i>qacEdelta.</i> Bleomycin and colistin resistance genes were detected as <i>ble</i> and <i>pmrB_R256G</i>, respectively. Comprehensive analysis of MDR strains provided by WGS detected variable antimicrobial resistance genes and their precise resistance mechanism. WGS is essential for control and prevention strategies to combat the growing threat of AMR and the implementation of multifaceted interventions are needed.
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