Outbreak of Cholera still remains major public health problem in most of the developing countries including Nepal. A prospective study was carried out at Dhulikhel Hospital, Kathmandu University Teaching Hospital, Kavrepalanchok during 1st May 2004 to 31st October 2004. A total of 148 stool samples from patients with acute diarrhea were collected and further investigated for Cholera. The study was conducted to establish the causes of the outbreak of acute diarrheal disease, antimicrobial profiles of the stool isolates and parasitic co –infection in Cholera cases. The samples were subjected to standard recommended microbial procedures and confirmation of the isolates was done by seroagglutination using V.cholerae polyvalent O1 and 0139 antisera and monovalent Ogawa and Inaba antisera. Out of the 148 stool samples, 46 cases (31%) were found to be positive for V.cholerae serogroup O1, biotype ElTor, serotype Ogawa. Both sexes were equally affected. Young age group of less than 30 years were mostly affected. Brahmin was the most affected ethnic group. The isolates were sensitive to all the antibiotics tested except co-trimoxazole. Among the laboratory confirmed cholera cases 30% exhibited co-infection with other parasites among which Giardia lamblia and Ascaris lumbricoides were the most common.