Introduction: In world HIV-2 virus was first isolated in West Africa in 1986 and is mainly restricted to West Africa. Due to increase in population mobility HIV-2 has spread from West Africa to other parts of world. In India HIV-2 cases are comparatively less and having geographically diverse spread in various states. HIV-2 infection is generally characterized by a longer asymptomatic stage, lower plasma viral loads, and lower mortality rate than HIV-1 infection. Aims & Objective: This study was conducted to determine the seroprevalence of HIV-2 and dual infection in HIV-infected individuals at the tertiary care center in Maharashtra. Materials and Methods: All the samples submitted for HIV testing at ICTC in GMC Akola during 5 years study period were included in the study. Following NACO guidelines, after informed consent and pre-test counseling patient’s blood sample were tested for HIV using testing strategy III (7). The samples positive for HIV-2 or dual infection were sent to NARI, Pune for further testing and confirmation. Observations & Results Out of 67969 patient’s samples tested for HIV 1.97% (n=1341) were positive for HIV antibodies. Out of 1341 patients 58.9% were male and 41.1% were female patients. Out of total samples tested 1.95% (n=1329) samples were HIV-1 positive, 0.014 % (n=10) were HIV-2 positive and 0.0029 % (n=2) were positive for both HIV-1 and HIV-2 (dual infection). Compared with HIV-1, HIV-2 positive patients represent very small proportion of all diagnosed HIV patients. Out of 1341 positive patients 58.9% (790) were male patients and 41.1% ((551)) were female patients. From all HIV-2 positive patients 50% patients were in age group of 31-40 years. Conclusion: In present study we conclude that the incidence of HIV-2 and dual infection does occur in Akola but it is comparatively less prevalent than other parts of India. As HIV-2 may pose serious threat in future, enhancing the diagnosis by detecting HIV-2 by various methods and correct treatment of HIV-2 infection according to new guidelines is the need of time for India.