Lymphadenopathy is one of the leading and persistent signs during the progression of human immunodeficiency virus (HIV) infection. Lymphadenopathy, as a clinical sign, can become one of the important guiding tools for detecting hidden HIV-positive cases. The present study was conducted to assess the incidence of HIV positivity in previously undiagnosed patients of HIV presenting with lymphadenopathy, comparing it with the current trends of HIV infection in the community, and to ascertain the etiology of lymphadenopathy in HIV-positive and HIV-negative cases. Over a period of 6 years, a total of 1082 cases of extra-inguinal lymphadenopathy were screened for HIV infection. Simultaneously, fine needle aspiration cytology of the enlarged lymph nodes was performed to know the etiology. The incidence of HIV positivity was found to be 2.3% among the patients presenting with lymphadenopathy. Among the HIV-positive cases, tuberculous lymphadenopathy was found in 60% of cases as compared to 28% of cases in HIV-negative cases. Alarmingly high incidence of HIV infection among the lymphadenopathy cases has been observed. This necessitates the importance of exercising vigilance, caution, and a high degree of suspicion while dealing with patients of lymphadenopathy, to protect ourselves as well as a much larger population at risk from this scourge.