Introduction: The outbreak of COVID-19 has spread worldwide. The evidence about risk factors of healthcare workers who infected COVID-19 is limited. This study aims to describe characteristics and influencing factors of the COVID-19 infection in healthcare workers. Methodology: The study was performed among COVID-19 infected and uninfected healthcare workers in three hospitals in Wuhan. A total of 325 healthcare workers participated; among them 151 COVID-19-infected healthcare workers were included. Characteristics of infected healthcare workers, and influencing factors including exposure histories, the use of protective equipment in different risk conditions and areas, perceptions, emotions, satisfactions and educations were described and analyzed. Results: Healthcare workers got infected clustered mostly in the physical examination center. When performing general operations on confirmed or suspected patients, the use of protective equipment including the effectiveness of masks (p < 0.001), gloves (p < 0.001); and the use of gloves (p < 0.001), suits (p < 0.001), gowns (p < 0.001), shoe covers (p < 0.001), and hats (p < 0.001) were protective factors. The use of protective equipment was a protective factor in most cases. Negative emotions and dissatisfaction to the hospital response were associated with the increased risk of infection. Conclusions: The use of protective equipment, emotions and satisfactions to hospital responses are key COVID-19-infected factors. The awareness, the supply and the use of protective equipment, the layout of departments and other environmental and management factors should be strictly equipped. In addition, hospitals should also pay attention to emotions and satisfaction of healthcare workers.