BACKGROUND: The serine carbapenemase enzymes (KPC) which produce from bacteria klebsiella pneumoniae today have been emerged as one of the β-lactamase enzymes that is capable to inactivating the last line of carbapenems. The gene encoding the K. pneumonia (blaKPC) belongs to gene carried on plasmid among Enterobacteriaceae family, which has modulation for the infections control so this study is aimed to spot the presence and evaluate blaKPC gene expression by real-time PCR in local isolates of K. pneumonia. METHODS: Forty-seven of K. pneumonia isolates were isolated from different clinical samples (blood, sputum, urine, wounds and burns) from patients in separate hospitals in Baghdad., Antimicrobial sensitivity test was carried out by vitik-2 system and Kirby- Bauer method. The PCR was employed to detect carbapenemase gene. RESULTS: The results of this study showed that all explored isolates were resistant to Ertapenem, Meropenem and imipenem 47(100%). Phenotypically, all the isolates had carbapenemase which hydrolyzed the carbapenem antibiotics. Furthermore, the isolates showed (100%) resistance to Cefazolin, Ampicillin and Amoxicillin/ Clavulic acid. However, the most effective antibiotic was Levofloxacin (91.5%). The results of conventional PCR technique for the detection of blaKPC gene showed that 38 (80.9%) isolates of carbapenem-resistant K. pneumoniae harboured blaKPC gene (1010 bp), while none carried other carbapenemase genes including blaNDM1, blaVIM and blaIMP genes. High levels of carbapenem resistance was clarified by the imipenem and meropenem MICs determination. All 38 isolates were positive in CNPT. Furthermore, the 38 isolates showed over expression of blaKPC gene compared with housekeeping rpo gene in Real-Time PCR. CONCLUSIONS: According to these results, the resistant isolates to carbapenem were belong to the present and high level expression of blaKPC gene in our local isolates.
BACKGROUND: The serine carbapenemase enzymes (KPC) which produce from bacteria klebsiella pneumoniae today have been emerged as one of the β-lactamase enzymes that is capable to inactivating the last line of carbapenems. The gene encoding the K. pneumonia (blaKPC) belongs to gene carried on plasmid among Enterobacteriaceae family, which has modulation for the infections control so this study is aimed to spot the presence and evaluate blaKPC gene expression by real-time PCR in local isolates of K. pneumonia. METHODS: Forty-seven of K. pneumonia isolates were isolated from different clinical samples (blood, sputum, urine, wounds and burns) from patients in separate hospitals in Baghdad., Antimicrobial sensitivity test was carried out by vitik-2 system and Kirby- Bauer method. The PCR was employed to detect carbapenemase gene. RESULTS: The results of this study showed that all explored isolates were resistant to Ertapenem, Meropenem and imipenem 47(100%). Phenotypically, all the isolates had carbapenemase which hydrolyzed the carbapenem antibiotics. Furthermore, the isolates showed (100%) resistance to Cefazolin, Ampicillin and Amoxicillin/ Clavulic acid. However, the most effective antibiotic was Levofloxacin (91.5%). The results of conventional PCR technique for the detection of blaKPC gene showed that 38 (80.9%) isolates of carbapenem-resistant K. pneumoniae harboured blaKPC gene (1010 bp), while none carried other carbapenemase genes including blaNDM1, blaVIM and blaIMP genes. High levels of carbapenem resistance was clarified by the imipenem and meropenem MICs determination. All 38 isolates were positive in CNPT. Furthermore, the 38 isolates showed over expression of blaKPC gene compared with housekeeping rpo gene in Real-Time PCR. CONCLUSIONS: According to these results, the resistant isolates to carbapenem were belong to the present and high level expression of blaKPC gene in our local isolates.
Authors: Ibtesam Ghadban Auda; Israa M S Al-Kadmy; Sawsan Mohammed Kareem; Aliaa Khyuon Lafta; Mustafa Hussein Obeid A'Affus; Ibrahim Abd Aloahd Khit; Abdulaziz Abdullah Al Kheraif; Darshan Devang Divakar; Ravikumar Ramakrishnaiah Journal: J AOAC Int Date: 2016-11-30 Impact factor: 1.913