To evaluate the awareness of HIV/AIDS prevention education, and the acceptance of HIV testing among residents on Likoma Island, Malawi, a cross-sectional, population-based study of 579 residents aged =15 years from seven villages on Likoma Island was conducted during July and August 2007. Most of the subjects studied could correctly answer questions about their awareness of AIDS and knowledge of the ways to reduce HIV transmission. Moreover, the proportion of respondents (65.8%) who possessed complete knowledge of HIV/AIDS prevention was greater than the national average. By contrast, condom utilization was slightly lower. Our results also showed that a high proportion of respondents (70.3%) had been HIV tested at any time, 93.5% of them voluntarily. Among correlated factors, females [adjusted odds ratio (AOR) = 1.7, 95% CI 1.1–1.6] and polygamous individuals (AOR = 3.3, 95% CI 1.5–7.0) were more likely to receive an HIV test. Past experience of being HIV tested was a strong predictor of possessing good knowledge and attitudes towards HIV/AIDS prevention. We conclude that antiretroviral treatment provided by Likoma District Hospital has led to the successful scale-up of HIV testing in Likoma Island and consequently improved the awareness of HIV/AIDS. However, the use of condoms remains largely unsupported, and there is therefore still a need to intensify general HIV/AIDS education on the island.