The objective of this study was to determine acceptability of sexual health discussion and testing among adolescents, parents, and health care providers (HCPs). We conducted a cross-sectional study of acute care patients (aged 14-19 years), parents, nurses, and physicians/nurse practitioners. Subjects rated sexual health discussion, sexually transmitted disease and pregnancy testing as acceptable/unacceptable and verbally explained their reasoning. Reasons were transcribed verbatim, coded, and analyzed. Agreement with individual adolescent responses was determined. Controlling for visit type, logistic regression was used to evaluate association between location and acceptance with odds ratios (ORs). Visits were categorized as reproductive/nonreproductive. Subjects included 127 adolescents, 90 parents, and 190 HCP responses. Mean adolescent age was 15.5 years; 57% were female; 43% reported previous intercourse; 43% were black, and 39% were white. Most visits (76%) were nonreproductive. Adolescents and parents were highly accepting of discussion and testing and were more accepting than HCPs. Acceptance was more likely for reproductive versus nonreproductive visits (P ≤ 0.05). Emergency department HCPs were more likely to accept discussion (OR, 2.8) and sexually transmitted disease testing (OR, 3.1) than urgent care HCPs. Acceptance themes among adolescents were comfort with system/HCP, confidentiality concerns, and desire for good health; among parents, acceptance themes were identification as a safe place for disclosure and desire for adolescent good health and information; and among HCPs, these were relevance to chief complaint and perception of appropriate care. In contrast to HCPs, adolescents and parents expressed broad support for sexual health discussion and testing, which may inform HCP practice to improve access to care.