Background: The total number of people with HIV (Human Immunodeficiency Virus) infection in India is estimated to be 10% of all global cases. People living with HIV in India often experience discrimination while receiving health care due to inadequate knowledge and fear among health care professionals. Data presented in this paper represents the first phase of a six-year study being conducted at a Medical College in Mumbai, India. Information from this phase of the study will be used to demonstrate the need for an HIV-specific training module for the first year medical students. Methods: A cross-sectional study was conducted among 200 first-year medical students in Mumbai, India to assess knowledge and attitudes as they relate to HIV infection. A self-administered survey was distributed among the medical students at a medical college in Mumbai. The survey sought student responses pertaining to knowledge of HIV risk and transmission, and attitudes towards HIV-infected people. Results: A response rate of 87% was obtained (174 out of 200). Overall, females showed less knowledge pertaining to issues related to human sexuality and HIV transmission when compared to their male peers. Anal intercourse was reported as a risk for HIV transmission by 3% of females as compared to 20% of males (p less than 0.05). Furthermore, 28% of females reported no relationship between the risk of contracting HIV and the type of sexual intercourse compared to 3% of males (p less than 0.05). In general, there were considerable misconceptions regarding the spread and risk of HIV transmission among all medical students. Sixty six percent (66%) of females were comfortable having HIV infected doctors and nurses (co-workers) in clinics and hospitals compared to 36% of males. Forty-four percent (44%) of the medical students preferred not being friends with HIV infected individuals. Sixty-two (62%) percent of the students favored abstinence only messages for prevention of HIV among teenagers. Discussion: Knowledge regarding risk and routes of HIV transmission was lacking among the medical students. Attitudes of the students toward HIV-infected individuals could be best described as ambivalent. However, female students showed more positive attitude towards HIV infected people than their male peers. Conclusions: Based on our findings we recommend the development and integration of a HIV training module in the first year medical curricula in order to address gaps in knowledge and provide training for the development of positive altitudes and tolerance toward HIV infected people. (Contains 2 tables.)