The Centers for Disease Control and Prevention recommends universal human immunodeficiency virus (HIV) screening in emergency departments (EDs) for patients (above 13 years). In 2017, Jackson Health System (Miami, FL) established a successful adult ED-based universal opt-out HIV screening program. However, when expanded to its affiliated pediatric ED (PED) as an opt-in protocol, only 1.3% of all adolescents received universal screening (only 1.7% were tested based on their chief complaints). We explore barriers to PED testing by comparing the attitudes of providers, patients, and caregivers. Institutional review board-approved surveys were administered anonymously to each group. Raw aggregate data were analyzed for trends. Providers' responses were stratified by role and responses were compared via a Mann-Whitney test. Twenty-four providers responded; they agreed that testing is warranted (x = 3.58 σ 1.32) and not time-consuming (x = 2.33 σ 1.05). There was no agreement on the perceived role of providers to perform testing. Attendings reported a high chance of breaching patients' confidentiality (x̄ = 3.27 σ 1.35), while nurses did not (x̄ = 2.33 σ 1.51). Of the 62 patient responses, 62.9% were amenable to testing, 69.4% would consent to 4th-generation serum testing if less invasive options (e.g., oral swabs) were unavailable, and 77.4% would inform caregivers of results. Of the 37 caregiver responses, 91.9% felt comfortable with their child consenting to testing. In this population, patients and caregivers appear amenable to universal screening. Low testing rates may be explained by providers' disagreement on their role to perform testing and concerns about confidentiality. Future studies should further explore protocol-based flaws and providers' views.