European Journal of Biomedical and Pharmaceutical Sciences

DETECTION AND CONFIRMATION OF PLASMID MEDIATED ANTIBIOTIC RESISTANCE AND RESTRICTION ENZYME ANALYSIS OF PLASMIDS EXTRACTED FROM ?-LACTAMASE PRODUCING UROPATHOGENS

Mobashshera Tariq and Aruna K*.

ABSTRACT

Extended spectrum β-lactamases (ESBL) are functionally defined as β-lactamases that are able to hydrolyze extended-spectrum cephalosporins (ceftazidime or cefotaxime) and monobactams (aztreonam) and, are inhibited by β-lactamase inhibitors, such as clavulanic acid, whereas they cannot hydrolyze cephamycins (cefoxitin or cefotetan) or carbapenems (imipenem or meropenem) efficiently. ESBLs can arise as a point mutation in blaTEM gene, however, a number of these enzymes are recently found to be encoded by plasmid mediated resistance mechanisms that allow increased spread of antibiotic resistance through efficient plasmid transfer. In the current study, plasmid mediated antibiotic resistance was confirmed by PCR amplification of resistant genes from extracted plasmids and plasmid loss studies carried out on 167 ESBL producing uropathogens. Restriction digestion analysis of plasmids from ESBL producer’s using EcoRI, BamHI and HindIII revealed similar band patterns in most of the E.coli and K.pneumoniae isolates indicating their clonal relatedness whereas P.aeruginosa revealed different band patterns indicating clonal un-relatedness and more than one source of origin for spread of antibiotic resistance. Conjugation and transformation studies revealed both intra as well as inter-species plasmid transfer indicating ease of transfer of antibiotic resistance. Our study hence gives an insight into the growing problem of antibiotic resistance and an urgent need for efficient control practices and prevention of such outbreaks.

Keywords: ESBL, EcoRI, BamHI, HindIII, PCR, uropathogens, Antibiotic Resistance.