Stigma has negatively influenced the lives of people living with HIV since the beginning of the epidemic. It affects every facet of their lives and can cause mental health problems, loss of human rights, and barriers to care. Studies in developing countries have shown a high prevalence of HIV stigma among health care workers. Few studies have been conducted in the United States. We used a validated instrument to survey 330 health care workers in Washington, DC, a high HIV prevalence area. The goal was to obtain data to assess the severity of the problem. We found that stigmatizing beliefs and attitudes were prevalent as reflected in responses from 66% of the participants. Of clinicians surveyed, 31% reported using double gloves. Participants with stigma training had lower stigma levels, whereas older individuals and support staff were more stigmatizing. Negative attitudes affect access to care and have major public health implications.