Listeria monocytogenes is responsible for causing the serious infectious disease listeriorosis which occurs due to food contaminated with Listeria. The study employed herein concerned with the isolation and identification of L. monocytogenes from foodstuffs and water. Hundred different food stuffs samples (milk, cheese and meat) and 50 fresh water samples were surveyed for presence of L. monocytogenes. These samples were inoculated into two enrichment media and cultivated on Oxford agar. Presumptively positive isolates were determined by morphology of colonies, gram stain, catalase test, haemolysis on sheep blood agar and motility test. The isolates were further identified to species level by 10300 API Listeria strips, PCR was done for all specimens to evaluate its accuracy. Out of 100 foodstuffs samples, 15 samples were showed positive results for Listria species. Out of 50 fresh water samples, 5 samples were showed positive results. Antibiotic susceptibility was done for all identified strains. All isolates were sensitive to penicillin G, ampicillin, tetracycline, amikacin and erythromycin.