Purpose: The present studied was performed in order to investigate the drug resistance of different non-lactose fermenting gram negative bacteria from clinical isolates. The bacteria were morphologically characterized through culturing and gram staining techniques were used for the identification of different bacterial strains. Methods: A total of 324 samples were collected from patients, after they were diagnosed by physicians at different hospitals at district Peshawar. Samples were morphologically identified by blood agar, MacConkey agar and Eosine Methylene Blue, identified by gram staining techniques. Modified Kirby-Bauer Disc diffusion method was used to test the in-vitro susceptibility of the identified isolates to different antibiotics. Results: The non-lactose fermenting gram negative bacteria were isolated from samples of blood (33.30%), pus/ wound (33.30%), urine (23.30%) and from ascetic/pleural fluids (10.20%). The study revealed that Pseudomonas aeroginosa showed high resistance against Gentamicin (74%) and Aztreonam (74%), followed by Ciprofloxacin (59.20%) and Amikacin (33.30). Tazocin was active as low resistance (18.50%) is shown. More resistance was seen in Morganella morganii against Aztreonam (77.7%) followed by Gentamicin (62.90%), Ciprofloxacin (40.70%). Tazocin show low resistance (3.70%). Multidrug resistant Proteus mirabillis was highly resistance to Gentamicin (66.60%), followed by Aztreonam (62.90%), Amikacin (55.50%), Ciprofloxacin (40.20%) and low resistance to Tazocin was (22.20%). Salmonella typhi demonstrated high resistance against Amikacin (62.90%), followed by Aztreonam (48.10%), Tazocin (40.70%). Gentamicin showed low resistance (29.60%), and hence it is more active against S. typhi. Conclusions: It can be concluded from the present study that different species of non-lactose fermenting gram negative bacteria have shown a different resistivity pattern. This study is a gate way for better and suitable management strategy for the infections caused by non-Lactose fermenting bacteria in the sampling region.