Background: Healthcare workers (HCWs) are continually exposed to hazards from contact with blood and body fluids of patients in the healthcare setting. Aim: To determine the prevalence of needlestick injuries (NSIs) and associated factors among HCWs in the Accident and Emergency Department of the University of Benin Teaching Hospital (UBTH), Benin City, Nigeria. Subjects and Methods: This was a crossâsectional study. Data were collected using a structured, selfâadministered questionnaire and analyzed using IBM SPSS version 20. Univariate, bivariate, and binary logistic regression analyses were done. The level of significance was set at P < 0.05. Results: The prevalence of NSIs 12 months preceding the study was 51.0% (50/98). Doctors 8/10 (80.0 %) and nurses 28/40 (70.0%) had the highest occurrence. Recapping of needles 19/50 (38.0%) and patient aggression 13/50 (26.0%) were responsible for most injuries. The majority 31/50 (62.0%) of the injuries were not reported. The uptake of postexposure prophylaxis (PEP) was low 11/50 (22.0%). The factors that were significantly associated with NSI include age 30 years and above (odds ratio [OR] =0.28, confidence interval [CI] = 0.11–0.70), work duration of three years and above (OR = 0.29, CI = 0.11–0.75), and being a nurse (OR = 3.38, CI = 1.49–9.93) or a paramedic (OR = 0.18, CI = 0.06–0.52). Conclusion: The high prevalence of NSIs among the HCWs, especially in doctors and nurses is an indication that HCWs in UBTH are at great risk of contracting bloodâborne infections. Efforts should be made to ensure that injuries are reported and appropriate PEP undertaken following NSIs.