Isolation and molecular identification of extended spectrum beta-lactamase producing bacteria from urinary tract infection

Abstract

Background: Recent years, the treatment of multi-drug resistant bacteria and their effect are very difficult due to the virulence factors modification. Based on the world wide thread, we have tried to identify the ESBLs producing bacteria from urinary tract infection patients. In result, the highly antibiotic resistant effect of Pseudomonas aeruginosa and Klebsiella pneumoniae were identified. Methods: Initially, Hexa disc diffusion method was performed to detect the multi-drug resistant bacteria using respective antibiotics of HX066, HX033 and HX077, HX012 discs. Consecutively, the ESBL producing ability of confirmed multi drug resistant bacteria was performed to detect their ESBL producing ability using specific extended spectrum beta lactamase (ESBLs) detection discs of Hexa G-minus 24. Furthermore, the ESBL producing ability of the bacteria was confirmed by ESBLs detection Ezy MIC™ E-test stripe method. Results and conclusions: In result, 10, 5 and 4 mm and 10, 14 and 8 mm zone of inhibition against imipenem (IPM), Ticarcillin/Clavulanic acid (TCC), Cefoperazone (CPZ) and Ampicillin (AMP), Norfloxacin (NX), Nalidixic acid (NA) antibiotics for P. aeruginosa and 16, 22 and 10, 18 mm zone of inhibition against ceftazidime (CAZ), methicillin (MET), ampicillin amoxyclav (AMC), co-trimoxazole (COT) of the HX077 HX012 for K. pneumoniae were observed. Based on the Clinical & Laboratory Standards Institute (CLSI) guidelines, both the bacteria were more resistant to tested antibiotics and it could be developed more resistant against all the tested antibiotics. In addition, the phenotypic detection of ESBL production effect was also performed against both the selected uropathogens, and the results were shown ≥22 mm, ≥27 zone of inhibition against all the tested antibiotics. Further, the genetic identification of multi plux PCR result was shown TEM, SHV and CTX-m genes were present in both the selected uropathogens. Finally, our results were correlated each other and concluded that the selected uropathogens were multi drug resistant effect and also ESBLs producer.

Keywords

• Antibiotic resistance
• Urinary tract infection
• Gram negative bacteria
• Phenotypic assay
• Gene identification
• CLSI guidelines