MOLECULAR DETECTION OF CO-PRODUCTION OF ESBL, AMPC AND INTEGRONS AMONG UROPATHOGENS IN A STUDY FROM MUMBAI
Mobashshera Tariq and Aruna K.*
ABSTRACT
Growing incidences of Extended Spectrum β-lactamases (ESBL) producing pathogens are a global issue. They are enzymes capable of hydrolyzing broad spectrum β-lactam antibiotics. In the current study, Phenotypic screening of ESBL producers was done from 747 gram negative uropathogens, among which 167 ESBL producers were identified. Screening of ESBL producers was done with the help of Antimicrobial Susceptibility Testing, Phenotypic Confirmatory Disc Diffusion Test, Double Disc Synergy Test and E- Test. Determination of minimum inhibitory concentration of 3rd generation cephalosporins, highlighted 37 ESBL producers to be either sensitive, or exhibit intermediate resistance to ceftazidime, ceftriaxone or cefotaxime. This indicates difficulty in routine ESBL detection where MIC is generally carried out as a screening step for ESBL detection. All ESBL producers showed presence of plasmid of approximately 50-60 kb in size. Confirmation of ESBL production was done by PCR amplification which showed presence of blaTEM (742bp) gene in 156 out of 167 ESBL producers, blaCTX-M (413bp) gene in 63/167 and blaAmpC (346bp) gene in 108/167 ESBL producers. Co-production of multiple β-lactamase genes was observed in 144/167 isolates. Class 1 and 2 integrons were detected in 51/167 (30.53%) and 11/167(7.78%) isolates respectively. Hence our study highlighted high prevalence of co-production of β-lactamases among uropathogens and the inefficiency of MIC as a phenotypic detection method in screening of β-lactamases.
Keywords: ESBL, PCDDT, DDST, PCR, uropathogens, Antibiotic Resistance.
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