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Diabetes mellitus is a chronic disorder and affects large segment of population and is a major public health problem. The infection leads to the early development of complication even after a trivial trauma, the disease progresses and becomes refractory to antibacterial therapy. Early diagnosis of microbial infections and screening for mechanism of drug resistance is aimed to institute the appropriate antibacterial therapy and to avoid further complications The aim of the present study is to find the prevalence of -lactamases mediated resistance among Gram negative bacteria isolated from urinary tract infection from diabetic patients. A prospective study was carried out on 1560 diabetic patients with urinary tract infection during the period of July 2011 to June 2015. 277 Gram negative bacterial were isolated and identified by standard laboratory techniques and screened for the presence of extended spectrum beta lactamase, AmpC lactamase, Metallo beta lactamase and confirmed by the respective confirmatory tests. 44.4 % of Gram negative bacilli were ES L producers. E.coli (34.1%) was the predominant ES L producer followed by Klebsiella pneumonia (30.9%). 6.5% of Gram negative bacilli were Amp C producers and Amp C production was seen only in Klebsiella pneumonia, Pseudomonas aeruginosa and Proteus mirabilis. 17.7% of Gram negative bacilli were M L producers, E.coli was the predominant M L producer (36.4%) followed by Acinetobacter baumannii, Klebsiella pneumoniae and Citrobacter freundii -lactamase producers are emerging threat and cause of concern for the clinicians, as it results in the resistance to penicillin, cephalosporins and limits therapeutic options. Screening techniques should be performed routinely to detect these -lactamase producers so that suitable antimicrobial therapy can be instituted.