Background: Brucellosis is endemic in many parts of the world including India and the disease is also the one of the important causes of PUO(Pyrexia of Unknown Origin).The population exposed to infected or contaminated animal and animal products are at a greater risk .The present study was conducted with an aim to determine the prevalence of Human Brucellosis in PUO cases among occupationally exposed individuals. Method: In the present hospital based cross-sectional study which was conducted in a tertiary care medical teaching institute in northern India. A total of n=369 samples of serum from PUO cases were collected and tested for Human Brucellosis by RBPT, STAT, iELISA. Result: Overall seropositivity in our study by different tests was found to be 0.5%, 0.8%, and 1.6% by RBPT,STAT and iELISA respectively. Conclusion: Highest seropositivity was found to be in Nomads (36.3%) followed by Farmers (27.2%).With respect to the symptoms generalized aches (90.8%) was most common followed by anorexia(64.4%).Also in our study of all the serum samples collected 20.3% gave history of consumption of Gau Mutra of which 2.6% of the samples were found to be seropositive. However inspite of the low prevalence of Human Brucellosis, clinicians must keep the index of suspicion high of the possibility of a patient suffering from Brucellosis while encountering a case of PUO and it is adisable to elicit a history of certain occupation and risk factor for Brucellosis in patients with PUO.