| Summary: | Seronegative women are susceptible to primary rubella virus (RV) infection during pregnancy, which can cause fetal damage. Vaccination represents the main strategy in rubella prevention. The aim of this study was to analyze changes in the rubella seroprevalence and identify populations with a high susceptibility to RV. A cross-sectional study was performed on 6914 Caucasian fertile women who had <i>Toxoplasma gondii</i>, other viruses, Rubella, Cytomegalovirus, and the herpes simplex virus (TORCH) screening in two distinct periods—1452 at the Timișoara Municipal Hospital, Romania (Group 1: 2008–2010) and 5462 at the laboratory Bioclinica S.A., Timișoara, Romania (Group 2: 2015–2018). The RV seroprevalence decreased (Group 1 versus Group 2; 94.1% (92.7–95.2) versus 91.4% (90.6–92.1), OR = 0.76 (<i>p </i>= 0.0007)). According to the year of birth and eligibility to vaccination program, RV seroprevalence rates were 82.4% (76.8–86.8)/1997–2004, 85.4% (80.5–89.3)/1995–1996, 90.1% (89.0–91.1)/<1989, and 95.8% (94.7–96.6)/1989–1994. No significant difference in the RV seropositivity according to the place of residence was found. The overall RV susceptibility increased from 2008–2010 to 2015–2018. The highest susceptibility was found in women born between 1997–2004 eligible for measles-mumps-rubella (MMR) vaccine through the family practice system and the lowest in women born between 1989–1994 eligible for monovalent rubella vaccine conducted in schools.
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