Background: Conflicting reports exist regarding the HIV-1 infection on the risk of malaria. A transient almost one-log elevation in HIV viral load occurs during febrile malaria episodes; in addition malaria is enhanced in HIV infected patients. HIV and malaria co-infection might also have facilitated the geographic expansion of malaria in HIV prevalent areas. We prospectively studied malaria patients for HIV infection from Mumbai. Materials and Methods: A total of 171 malaria patients and 28749 normal voluntary blood donors studied for their HIV status. On microscopical examination of blood, all patients showed a ring form of P.falciparum. Blood screened for HIV-1&2 antibodies by micro well ELISA using Enzaids & J Mitra kits followed by confirmation using Western Blot (Innogenetics, Belgium) analysis. Results: Thirteen malaria patients (7.6%; Odds ratio= 4.45; P value <0.0001) and 521 blood bank donors found to be HIV reactive. Among the 13 HIV reactive patients, we found that eight patients (4.6%) who were Elisa borderline reactive and western blot positive (p24) due to cross-reactive antibodies. Five Malaria patients (2.9%) found to be HIV-1 positive by ELISA and Western Blot confirming co-infection. Conclusion: It seems that our findings suggest that HIV-1 and Malaria co-infection is associated with severe malaria in hyper endemic countries like India.