In Italy, the inclusion of measles vaccine in children immunization schedule and the promotion of national mass vaccination campaigns increased measles vaccination coverage. However, measles outbreaks continue to occur increasingly involving adolescents and adults. The aim of this study was to evaluate the prevalence to measles antibody in a sample of Italian population between 1993 and 2018.Human serum samples from subjects aged 3–40 years were collected between 1993 and 2018 and tested for measles IgG antibodies by commercial ELISA.During the study period, the 3–10-year-old age group showed the most important change, with a significant increase in 2003–2007 in both seroprevalence and IgG levels, followed by a slow decrease. The 11–18-year-old age group showed relatively stable seroprevalence rates and IgG levels over the years. The 19–30-year-old group showed stable seroprevalence rates, albeit with a decrease in IgG levels. After a significant increase in 1999–2002, the 31–40-year-old age group had high seroprevalence rates and IgG levels.Despite efforts at national level for implementing measles vaccination, a large proportion of the population is still susceptible to measles. Even if vaccination coverage increases enough to achieve the level of immunization required for herd immunity in new birth cohorts, outbreaks will continue to occur if there are immunity gaps in older age groups. Establishing policies for measles vaccination targeting adult population is needed to close immunity gaps and reach the elimination goal.