<p>The First Egyptian Report Showing the Co-Existence of <em>bla</em>_NDM-25, <em>bla</em>_OXA-23, <em>bla</em>_OXA-181, and <em>bla</em>_GES-1 Among Carbapenem-Resistant <em>K. pneumoniae</em> Clinical Isolates Genotyped by BOX-PCR</p>

Abstract

Background and Objective: The emergence of carbapenem-resistant K. pneumoniae (CRKP) continues to escalate and is alarming because of the emergence of pan drugresistant strains. The objective of this study was to investigate the existence of 12 carbapenemase genes among CRKP clinical isolates. Methods: Ninety-six Klebsiella spp. clinical isolates were collected. The isolates were identified phenotypically and genotypically. These isolates were screened for susceptibility to 24 different antibiotics. The modified Hodge test (MHT) and the Carba Nordmann/Poirel (NP) test were used to phenotypically screen carbapenem-resistant strains for carbapenemase production. Phenotypic characterization of carbapenemases was performed using the combined disk synergy test (CDST). Additionally, the presence of 12 carbapenemase genes in CRKP isolates was investigated. The DNA sequence of bla NDM and bla GES genes was determined. The BOX-PCR technique was used to determine the clonal relationship between CRKP isolates. Results: All carbapenem-resistant isolates were related to K. pneumoniae. Susceptibility testing showed that 19.79% (19/96) of the collected isolates were carbapenem-resistant. Of the CRKP isolates, 68.42% (13/19) tested positive for the MHT and Carba NP test. CDST showed that 42.11% (8/19), 63.16% (12/19), 47.37% (9/19), and 73.68% (14/19) of the CRKP isolates tested positive for the inhibitory effect of clavulanic acid, sulbactam, phenylboronic acid, and tazobactam, respectively, while 84.21% (16/19) and 68.42% (13/16) tested positive for the inhibitory effect of EDTA and mercaptopropionic acid, respectively. It was found that 10.53% (2/19) of the isolates tested positive for the inhibitory effect of sodium chloride. Molecular investigation of carbapenemases showed that 26.32% (5/19), 73.68% (14/19), 21.05% (4/19), 10.53% (2/19), and 5.26% (1/19) of the isolates tested positive for bla NDM , bla OXA-48 , bla OXA-181 , bla OXA-51 , and bla OXA-23 , respectively. None of the isolates tested positive for bla OXA-40 and bla OXA-58. Two allelic variants of bla NDM (bla NDM-1 and bla NDM-25) were detected. BOX-PCR revealed high clonal relatedness between CRKP isolates. Conclusion: MHT was more sensitive than Carba NP test for evaluating carbapenemase production and class D carbapenemase genes were the most prevalent of the 12 carbapenemase genes that were evaluated.

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

“… 46 Although there is a marked increase in the resistance of amikacin, colistin, and tigecycline in recent studies in Egypt, they are still effective against carbapenem-resistant Enterobacteriaceae. 37 , 39 , 47 Molecular typing is a useful tool for determining the genetic relatedness of bacteria. This is particularly important in the investigation of the epidemiology of several bacterial infectious diseases accompanied by HAI.…”

“…Similar to our study, recent two studies were conducted also in Egypt that reported the prevalence of diverse clones from CRKP using the PFGE and BOX PCR as molecular typing methods, respectively. 37 , 49 …”

“…35 As reported in other previous studies in Egypt, our study also concluded that the predominant species among the isolated CRE was MDR K. pneumoniae, which represents a frightening causative agent of different types of infections among hospitalized pediatric cancer patients. [35][36][37][38] It was reported that class D carbapenemases (bla OXA-48 = 58%) are predominant in K. pneumoniae followed by the bla NDM-1 gene, in several studies conducted in different countries notably in the Mediterranean region including Spain, France, Turkey, Tunisia and Egypt. 20,21 Furthermore, the majority (41.8%) of the K. pneumoniae isolates in the current study harbored one or more carbapenem resistance determinants (Figure 2) that agreed with previously reported studies.…”

Expanding of ST11 Carbapenemase-Producing Klebsiella pneumoniae Subclones in a Chinese Hospital, Shenzhen, China

“…The existence of OXA-23 among CRKP clinical isolates in Egypt was also reported recently. 23 This suggests that we should be alert to the spread of OXA-23 in K. pneumoniae , which is only prevalent in Acinetobacter baumannii . 23 However, MLST schemes corresponding to CPKP strains have remained imprecisely defined.…”

“… 23 This suggests that we should be alert to the spread of OXA-23 in K. pneumoniae , which is only prevalent in Acinetobacter baumannii . 23 However, MLST schemes corresponding to CPKP strains have remained imprecisely defined. 9 , 10 cgMLST analysis may eliminate spurious MLST associations.…”