Despite the Centers for Disease Control and Prevention recommendations to test patients ages 13 to 64 years for HIV at health care settings, routine HIV testing is lacking. As a result, many people are unaware of their HIV seropositive status. The purpose of this quantitative cross-sectional study was to examine relationships between HIV testing and provider type, knowledge, attitudes, and behaviors regarding HIV testing in the acute care setting. The study was informed by social cognitive theory. Using a convenient sampling method, a questionnaire derived from previous surveys (Society of General Internal Medicine and University of Washington) was sent to 600 eligible acute care providers from a suburban Chicago hospital who treated HIV-negative patients ages 13 to 64 years. Completed surveys were received from 88 participants. Chi-square and multiple logistic regression testing showed no significant relationships between HIV testing and provider type (p = .09), age (p = .91), gender (p = .84), experience (p = 1), and race/ethnicity. However, knowledge of HIV testing regulations and positive attitudes about HIV testing were significantly associated with the likelihood of offering an HIV test (p = .026, p = .004 respectively). Results have some clinical importance, but also indicated a lack of routine opt-out HIV testing. Results may be used to promote HIV testing among acute care providers which could reduce HIV-status unawareness in the population.