Medical Laboratory science علوم المختبرات الطبية

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Browsing Medical Laboratory science علوم المختبرات الطبية by Author "Aseel Khaled Jamal ALSharif"

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    Detection of Carbapenem- Resistant Genes in Klebsiella pneumoniae from Varios Clinical Samples.
    (Al-Quds University, 2025-08-09) Aseel Khaled Jamal ALSharif; اسيل خالد جمال الشريف
    Background: Klebsiella pneumoniae (K. pneumoniae) is a significant cause of hospital-acquired infections. Penicillin, cephalosporins, and carbapenems are the β-lactam antibiotics that most frequently prescribed to treat this type of infection. However, this bacterium has become multidrug-resistant (MDR), and it is clear that rise resistance around the world lead to threaten public health on global scale. Carbapenem is often considered the last effective treatment option for multidrug-resistant gram-negative bacteria. Unfortunately, K. pneumoniae has developed resistance to the last effective beta-lactam antibiotic which is carbapenem. This resistance primarily results from the production of carbapenemases, such as K. pneumoniae Carbapenemase, KPC; Verona Integron Metallo Beta-lactamase, VIM; New Delhi Metallo-β-Lactamase-Mediated Carbapenem, NDM; and Oxacillin-Hydrolyzing Carbapenemases, OXA-48, which can be acquired in both hospital and community settings. Aims: K. pneumoniae strains resistant to carbapenem (CR-Kp) are major health care-associated pathogens found globally. This study aimed to reveal the antibiotic resistance of clinical CR-Kp isolates and to find out whether carbapenemase genes, such as bla KPC, bla VIM, bla NDM, and bla OXA-48, are present, using multiplex PCR techniques. Methodology: A total of 100 bacterial isolates were collected from Prince Alia Governmental Hospital in Hebron, Palestine. The isolates were identified as K. pneumoniae based on their colony morphology and subsequently confirmed using the VITEK 2 Compact system. Antibiotic susceptibility testing (AST) was determined using the same system. The ESBL was conducted by the double disk synergy test (DDTS). The screening for carbapenem resistance was carried out using meropenem (MEM) disks, followed by multiplex PCR to detect the presence of the carbapenemase genes bla KPC, bla VIM, bla NDM, and bla OXA-48 Result: Out of the 100 K. pneumoniae isolates included in this study, analysis indicated that 77 of the isolates (77%) showed extended-spectrum beta-lactamase (ESBL). Out of 100 isolates 35 representing (35%) were carbapenem resistant (CR). All of the 35 isolates were tested for the presence of carbapenemase genes. The results for the genes of carbapenemase were as follows: The bla OXA-48 gene was found in 32 of the 35 samples (91.4%); the bla NDM gene was in 29 of the 35 samples (82.8%); and the bla KPC gene was in 2 of the 35 samples (5.7%). However, the bla VIM gene was not found in any of the samples (0.0%). The co-occurrence of both bla NDM and bla OXA was observed in 26 out of 30 samples (74%). Conversely, the co-occurrence of bla KPC and bla OXA was found in only one isolate (28%). Additionally, fifteen isolates that tested positive for extended-spectrum beta-lactamases (ESBL) but were sensitive to the meropenem (MEM) disk (carbapenem sensitive) were randomly chosen to be checked for carbapenemase genes. Out of the 15 isolates, blaVIM was found in 7 out of 15 samples (46.6%); while the bla OXA-48 gene was found in 4 out of 15 samples (26.6%). In contrast, the bla NDM gene was detected in only one sample (6.7%). Conversely, the co-occurrence of bla VIM and bla OXA was found in 4 out of 15 samples (26.6%). Conclusion: The study findings clearly indicated that the rate of carbapenem-resistant Enterobacteriaceae (CRE) produced by K. pneumoniae isolates was high, making it a significant cause of healthcare-associated infections. Resistance to carbapenems is not limited to carbapenem-resistant isolates; some strains exhibiting this resistance can also be found among extended-spectrum beta-lactamase (ESBL) isolates. To ensure effective treatment, it is important to keep track of the local molecular epidemiology of CRE-resistant genes, and/or another reliable assay. Keywords: K.pneumoniae, Carbapenemases gene, multiplex PCR, Antibiotic susceptibility. blaKPC, blaVIM, blaNDM, and bla OXA-48.