Abstract Background Poor hand hygiene (HH) among health care workers (HCWs) in low-resource healthcare settings has continued to increase the spread of infectious diseases, including COVID-19. The present study aimed to assess the knowledge, attitude, adherence, and barriers to HH practices among HCWs during the COVID-19 pandemic in Nigeria. Methods The study was an online cross-sectional survey using a tailored questionnaire distributed through chain referral sampling among southern Nigerian HCWs during the first wave of the COVID-19 pandemic (February to August 2020). The primary outcomes were knowledge, attitude, adherence to HH guidelines, and the barriers limiting compliance with the guidelines. Data were analysed using percentage, frequency, mean, standard deviation, one-way ANOVA, and multiple linear regression. The study timeline was from 15 April to 31 July 2020. Results Four hundred and fifty-four HCWs (236 males and 218 females) participated in the study. The participants had a moderate knowledge of standard HH protocol (mean ± SD) 62.45 ± 10.82%, positive attitude 84.34 ± 11.32%, and high adherence to the standard guidelines 81.21 ± 9.49%. There was no significant difference in knowledge, attitude, and adherence across the healthcare professions. Increasing age (β = 0.186, p < 0.003) and knowledge (β = 0.229, p < 0.001), and decreasing negligence (β = − 0.178, p = 0.004), and forgetfulness (β = − 0.159, p = 0.012) were the significant predictors of effective HH practices. Conclusion Nigerian HCWs had moderate knowledge, a positive attitude, and adhered to the standard HH practices during the COVID-19 pandemic. However, the major barriers were institutional-based factors such as the inadequate supply of HH items, competing job demands, emergencies, increased workload, and personal characteristics such as age, poor knowledge, negligence, and forgetfulness. We recommend that hospital management provide their staff with adequate HH materials and continued infectious disease training.
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