BACKGROUND: Brucellosis is endemic to North Karnataka. It is an important public health and veterinary disease here. The clinical symptoms of human brucellosis are protean, nonspecific and it simulates several other febrile diseases. The disease, therefore, cannot be diagnosed on clinical grounds alone, and microbiological confirmation is a must for evidence based antibiotic therapy. Serology is the mainstay of diagnosis of brucellosis; however, it has a lot of limitations. Therefore, a better method for diagnosis especially in the endemic areas is a need. The aim of this study is to establish PCR on clinical brucella isolates using genus specific primers. AIMS: To standardize an in-house polymerase chain reaction (PCR) for identification of Brucella isolates using genus specific primers.