Healthcare workers (HCWs) are the backbone of the healthcare system and a skilled and healthy workforce is vital during a health crisis, such as the present coronavirus disease 2019 (COVID-19) pandemic. Healthcare workers are at higher risk of exposure to and transmission of the severe acute respiratory coronavirus-2 (SARS-CoV-2). Hence, HCWs should possess good knowledge and attitudes toward COVID-19 to protect not only themselves but also their colleagues, families and the larger community. Thus, the current study aims to assess the knowledge and practice of protective measures by HCWs in Saudi Arabia during the first wave of the COVID-19 pandemic to identify awareness of the disease, potential trends and associated predictors.