The current study was carried out to determine the sero-prevalence and molecular detection of Brucella species in persons involved in pork production in Punjab state of India. The sample size was selected using survey toolbox and a total of 123 blood samples were collected from pig farmers and slaughter house workers. The serum samples were tested using Rose Bengal Plate Test (RBPT), Standard Tube Agglutination test (STAT) and indirect ELISA. Polymerase chain reaction (PCR) on blood samples using B4 and B5 primers was carried out on all sero-positive and 20 randomly selected sero-negative samples. The Bruce ladder Multiplex PCR technique was further used for confi rming the species of Brucella positive samples. The results observed that 1 (0.81%) and 4 (3.25%) subjects were positive in RBPT/STAT and ELISA, respectively. Polymerase Chain Reaction confirmed the presence Brucella species in blood of one of the sero-positive samples and Bruce ladder multiplex PCR confi rmed the species to be B. abortus. Chi-square test was applied on the results to determine the signifi cant difference among various groups. A significant difference was found in the prevalence rates in different districts in Punjab. The results indicate that B. abortus is circulating in the persons involved in pork production in Punjab state of India. The preventive and control measures need to be enforced to prevent the occupational exposure of brucellosis to pig producers in Punjab (India).