Molecular Characterization of Carbapenemase-Producing Klebsiella pneumoniae Isolated from Egyptian Pediatric Cancer Patients Including a Strain with a Rare Gene-Combination of β-Lactamases

Abstract

Purpose Healthcare-associated infections caused by multi-drug-resistant (MDR) pathogens are a global threat. We aim to assess the clonal relatedness among carbapenemase-producing Klebsiella pneumoniae (CPKP) strains infecting Egyptian pediatric cancer patients. Materials and Methods Identification and antimicrobial susceptibility testing of 149 Gram-negative isolates obtained from pediatric cancer patients were performed by VITEK 2. Genes encoding carbapenemases and extended-spectrum β-lactamases were detected by PCR and verified by DNA sequencing of representative samples. The transferability of the plasmids harboring bla OXA-48 , from representative clinical samples, was evaluated by performing a conjugation experiment followed by PCR and MIC shift determination. Clonal relationships among the bla OXA-48 -harboring K. pneumoniae isolates were determined by enterobacterial repetitive intergenic consensus (ERIC)-PCR and pulsed-field gel electrophoresis (PFGE). Results Carbapenem resistance was observed in 59% of the isolates. The most prevalent species was K. pneumoniae (45.6%) and 57% of them were isolated from ICU. Fifty-nine % of the K. pneumoniae isolates were carbapenemase-producers and bla OXA-48 was detected in (58%) of them. One isolate co-harbored bla OXA-48, bla NDM-1 , and bla IMP-1 genes for the first time in Egypt. PCR and meropenem MIC shift confirmed the success of the transferability of representative plasmids to E. coli K12. ERIC and PFGE identified 93% and 100% of the K. pneumoniae with a similarity coefficient ≥85%, respectively, including strains with indistinguishable patterns, suggesting possible clonal dissemination. Conclusion Our findings underline the dissemination of diverse clones of MDR CPKP among Egyptian pediatric cancer patients. Hence, routine molecular characterizations followed by strict implementation of infection control measures are crucial to tackling this threat.

Phenotype-genotype correlations among carbapenem-resistant Enterobacterales recovered from four Egyptian hospitals with the report of SPM carbapenemase

“…Strikingly, carbapenem resistance was detected in 80.9% of the Enterobacterales isolates. This level of resistance is higher than recent studies in Egypt describing the isolation of CRE with the percentages 46% [ 26 ], 59% [ 27 ], 68.8% [ 28 ] from cancer patients and 48% from intensive care units [ 5 ]. In the study performed by Hassuna et al [ 29 ], very high frequency of carbapenem resistance (95%) was detected among the K. pneumoniae isolates.…”

Phenotypic and genotypic characterization of carbapenemases in carbapenem-resistant gram-negative bacilli isolated from adult cancer patients.

“…By using the disc diffusion method and the broth microdilution method, respectively, the results showed that 65 (60.2%) and 91 (84.3%) of the isolates were meropenem resistant [47]. This percentage of resistance (84.3%) exceeds the percentages reported in recent Egyptian research documenting the isolation of CRE, which were 46% [48], 68.8% [49] 59% [50], from patients with malignancy, and 48% from ICUs [51]. In the study conducted by Hassuna et al it was found that 95% of the K. pneumoniae isolates were resistant to carbapenem [52].…”

Genotyping and molecular investigation of plasmid-mediated carbapenem resistant clinical <i>Klebsiella pneumoniae</i> isolates in Egypt

“…All isolates were positive for the OXA gene, 56 (93%) were positive for the VIM gene, 51 (85%) were positive for the NDM gene, 38 (63%) were positive for the IMP gene, and 10 (17%) were positive for KPC gene. In Egypt, the OXA gene was reported as the most predominant carbapenemase with no or low prevalence of the KPC gene in many studies between 2020 and 2022 [37] – [39] . In contrast, Gandor et al .…”