Limited data exists on the persistence of neutralizing antibodies (NAbs) among SARS-CoV-2-infected healthcare workers (HCW). This study followed 26 HCW with mild COVID-19, assessing NAbs at three time points. A significant decrease in NAb titers was observed over time, correlating with a decline in systemic IgA antibody levels. This short-lived humoral protection highlights the need for continued infection prevention measures and potential periodic vaccination boosts for HCW.