This cross-sectional study describes Sexual Assault Nurse Examiner (SANE)/Forensic Nurse Examiner (FNE) program practices related to HIV testing, non-occupational post-exposure prophylaxis (nPEP), and common barriers to offering HIV testing and nPEP. A convenience sample of 174 SANE/FNE programs in the United States and Canada was drawn from the International Association of Forensic Nurses database, and program coordinators completed web-based surveys. Three quarters of programs had nPEP policies, 31% provided HIV testing; and 63% offered nPEP routinely or upon request. Using Chi-square and Fishers' exact tests a greater proportion of Canadian programs had an nPEP protocol (p = .010), provided HIV testing (p = .004), and offered nPEP (p = .0001) than U.S.-based programs. Program coordinators rated providing pre-/post-counseling and follow-up as the most important barrier to HIV testing and medication costs as the most important barrier to providing nPEP. Our results indicate HIV-related services are offered inconsistently across SANE/FNE programs.