The Development of a PCR-based method for the detection of Klebsiella pneumoniae Carbapenem resistance genes in clinical specimens
Keywords:
Carbapenem-resistant, Klebsiella pneumoniae, bla-OXA, bla-KPC, bla-NDMAbstract
Background: Extended-spectrum beta-lactamase-producing Klebsiella pneumoniae (ESBL-KP) has shown increasing resistance to carbapenems, which are among the last-resort antibiotics. Carbapenem-resistant K. pneumoniae (CRKP) poses significant challenges in healthcare settings due to its resistance to even potent antibiotics, making infections difficult to treat and control. CRKP is often associated with severe complications such as pneumonia, bloodstream infections, and urinary tract infections, particularly in immunocompromised patients.
Aim: This study emphasizes the need for specific identification protocols, individualized therapy, and effective infection prevention measures to control the spread of CRKP.
Materials and Methods: A total of 300 clinical specimens were collected from patients at Al-Hilla Teaching Hospital between April and September 2024, including blood (3), urine (2), wounds (10), and burns (10). Bacteria were isolated using standard media, identified using the VITEK-2 system, and subjected to antibiotic susceptibility testing according to CLSI guidelines (2024). PCR was employed to detect carbapenem resistance genes (blaKPC, blaNDM, blaOXA).
Results: Of the 300 specimens, 25 (8.33%) were identified as K. pneumoniae. PCR-based methods enabled rapid detection of resistance genes: blaKPC (96%), blaNDM (80%), and blaOXA (72%). Biochemical tests confirmed positive results for urease, methyl red, citrate, and indole tests. The VITEK-2 system facilitated extensive phenotypic and antibiotic susceptibility testing. Resistance patterns varied, with susceptibility highest to carbapenems and lowest to ampicillin (100% resistance). Molecular detection of the 16S rRNA gene achieved 100% accuracy.
Conclusion: The findings highlight the critical threat posed by CRKP and underscore the importance of molecular diagnostics in detecting resistance genes. This research contributes to understanding carbapenem resistance mechanisms, guiding the development of effective treatment strategies.
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Copyright (c) 2024 Alya Amer Rahi; Huda H. Al-Hasnawy, Batool Kereem Mohammed, Dina Tariq Sharara, Diana Jalal Albiaty
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