AbstractBackground: Pseudomonas species are the commonest pathogens causing nosocomial infections. Pseudomonas is basically resistant to many antibiotics and they are known to produce Extended Spectrum Beta Lactamase and Metalo beta lactamase. AIM: To detect Metalo beta lactamase producing Pseudomonas spp. from clinical samples in tertiary care hospital. Material and Methods: The study was agreed over a period of 6 months from January 2015 to June 2015. A total non repetitive of 329 Pseudomonas spp were isolated from unusual clinical samples like blood, pus and wound swabs, urine, body fluids, sputum, endo tracheal tube and secretions from the patients attending the hospital. Antimicrobial susceptibility test of all the isolates was performed by the disc-diffusion (Kirby Bauer disc diffusion method) according to CLSIs guidelines. All imipenem resistant isolates were tested for MBL production by Imipenem - EDTA double- disc synergy test (DDST) and Imipenem- EDTA combined disc test (CDT). Result: Of 329 samples, majority of the Pseudomonas spp were isolated from Pus/Wound 173 (52.58%) followed by Blood 103 (31.31%). The isolation rate was highest from pediatrics wards 112 (34.04%) and surgical wards 112 (34.04%). Total 329 samples, 24 (7.29%) isolates were MBL producer by CDST and DDST. Majority of the MBL producing Pseudomonas spp. were isolated from Pus/Wound 11 (45.83%) followed by Urine 10 (41.66%). The isolation rate was highest from Surgical wards 9 (37.5%) followed by the Pediatric wards 7 (29.16%). Conclusion: Detetion of Metallo beta lactamase isolates of Pseudomonas spp. will help to implement rational use of antibiotics and strictly adhere to the concept of “reserve drugs” are important to identify because it poses therapeutic problems and serious concern for infection control management. Keywords: Metalo Beta Lactamase (MBL); Pseudomonas Species; Nosocomial Infections.