HIV/AIDS is a significant public health issue for individuals living in the United States. Federal agencies and key stakeholders recommend routine HIV/AIDS screening among individuals aged 13-64 of age, yet few studies focus on the nurse practitioner's (NP) perspective. The purpose of this study was to examine routine HIV screening intentions, behaviors, and practices using a national sample of NPs. Cross-sectional study utilizing a random sample of 600 NPs from the American Association of Nurse Practitioners™ organization. A total of 141 NPs returned complete responses to the paper and pencil questionnaire and were included in the study. One-quarter (25%, n = 35) of the NPs reported routine HIV screening behavior. Half (48%, n = 67) reported intentions to increase routine HIV screening in the next year. HIV screening practices were reported as risk-based and patient-initiated among this sample of NPs. In the U.S. healthcare system, NPs serve as leaders in primary and secondary prevention, screening and identifying individuals who are unaware of their HIV infection. Greater coordinated efforts and new system approaches are needed to help NPs adapt HIV screening recommendations and practice guidelines.