Background: Standard precautions prevent the spread of infections in healthcare settings. Incompliance with infection control guidelines of healthcare workers (HCWs) may increase their risk of exposure to infectious disease, especially under pandemics. The purpose of this study was to assess the level of compliance with the infection prevention and control practices among HCWs in different healthcare settings and its relationship with their views on workplace infection control measures during the COVID-19 pandemic. Methods: Nurses in Hong Kong were invited to respond to a cross-sectional online survey, in which their views on workplace infection and prevention policy, compliance with standard precautions and self-reported health during pandemics were collected. Results: The respondents were dissatisfied with workplace infection and prevention policy in terms of comprehensiveness (62%), clarity (64%), timeliness (63%), and transparency (60%). For the protective behavior, the respondents did not fully comply with the standard precautions when they were involved in medical care. Their compliance was relatively low when having proper patient handling (54%) and performing invasive procedures (46%). A multivariate analysis model proved that the level of compliance of the standard precautions was positively associated with the satisfaction on infection control and prevention policy among high risk group (0.020; 95% CI: 0.005–0.036), while older respondents had higher level of compliance among the inpatient and outpatient groups (coefficient range: 0.065–0.076). The higher level of compliance was also significantly associated with working in designated team and having chronic condition of the respondents among high-risk and inpatient groups. Conclusions: Standard precautions are the most important elements to reduce cross-transmission among HCWs and patients while the satisfaction on infection control and prevention policy would increase the compliance among the high-risk group. An overall suboptimal compliance and poor views on the infection prevention and control guidelines is a warning signal to healthcare system especially during pandemics.