Introduction/Background: Cytomegalovirus (CMV) is the leading congenital infection, which can occur when women have a primary CMV infection, reinfection with a new CMV strain, or reactivation of a prior CMV infection during pregnancy. Due to exposure to young children, daycare workers (DCWs) may have an increased risk of CMV infection. Thus, we initiated a study to evaluate the risks and knowledge of CMV infection among DCWs in Quebec. Methods: We will recruit female DCWs (n=533) 18-45 years of age from daycare centers, as well as a control group (n=1659) of females without work-related exposure to young children. Participants will be followed for 12 months with blood and saliva collection, and surveys on relevant socio-demographic factors, knowledge, and behaviors. Conventional and strain-specific serology, will be used to detect primary and non-primary infections (i.e., reinfection or reactivation), respectively, and CMV shedding in saliva will be measured by PCR. Poisson regressions adjusted for clustering and weighted for differences in baseline characteristics will be used to calculate the relative risk of primary infection or reinfection, and frequency of shedding among DCWs compared to controls. Qualitative data gathered through semi-structured interviews will be analyzed using an inductive approach, to describe the knowledge and attitudes about CMV infection risks. Anticipated results: We hypothesize that CMV infection and reinfection with diverse viral strains are more frequent among DCWs than controls after controlling for potential confounders. We expect that DCWs will have a greater awareness of CMV and its potential severity than controls, and that most participants in both groups will be open to behavioral interventions to reduce the risk of (re)infection during pregnancy. Conclusions and implications for policy, practice or additional research: Our research will refine risk estimates and inform public health strategies to prevent congenital CMV infection through behavioral measures as well as vaccine development.