We consider the clinical presentation of patients with equivocal HIV test results in the ED (HIV Ab/Ag +, HIV Ab -) found through routine ED-based HIV screening and further describe those patients who presented to the ED during acute seroconversion (acute +) compared to the clinical characteristics of patients who were ultimately found to have false + initial screening results. We analyzed 4 years of HIV testing data (2016-2020) and determined the number of patients who had a Ab/Ag + screen. Patients with Ab/Ag + but a non-equivocal HIV+ lab signature (Ab/Ag +, Ab +, viral load > 0) were removed. Then we determined the remaining number with an equivocal laboratory signature (Ab/Ag +, Ab -). We separated those patients into 2 groups: false + (Ab/Ag +, Ab -, viral load 0) and acute + (Ab/Ag +, Ab -, viral load > 0). We conducted chart review on all patients with an equivocal laboratory signature and the clinical presentation was considered to detail patterns in false + compared to acute + patients presenting to the ED. We screened approximately 55,224 patients for HIV (16% volume) in 4 years. 787 patients had a Ab/Ag + result (1.4%) and, of those, 688 had non-equivocal positive HIV results (87.4% of Ab/Ag +, 1.2% of tested). 99 (12.5% of Ab/Ag +, 0.13% tested) were Ab/Ag +, Ab -. Of those 99, 73 had no detectable HIV RNA (false +, 9.3% of Ab/Ag +, 0.13% tested). 26 of the 99 with equivocal results had viral load > 0 (acute +, 3.3% of Ab/Ag +, 0.05% tested). Qualitative review of equivocal patient charts during the Ab/Ag reactive screening encounter showed statistically significance for acute positive results in younger male patients who have sex with men. 787 patients had a reactive screening test but 99 had an equivocal laboratory signature (12.5% of Ab/Ag+), making the information difficult to interpret during an ED encounter in high prevalence populations and challenging the ability to scale up ED based HIV screening, especially given the long turn around time for HIV RNA testing via PCR. ED based screening is an important strategy to help reach the WHO goal of eliminating HIV as a public health threat by 2030. However, the current algorithm and existing testing technology may not be best designed for acute clinical encounters and false + encounters are higher than previously reported. The results of this study detail characteristics of patients with equivocal test results that may improve clinical decision making in patients with false + compared to acute + laboratory signatures and suggest that young men who have sex with men and have a reactive HIV screening test in the ED should be considered HIV positive.