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Background Sero-positivity rates of the rubella virus among pregnant women vary widely throughout the world. In Tanzania, rubella vaccination is not included in the national immunization schedule and there is therefore no antenatal screening for this viral disease. So far, there are no reports on the sero-prevalence of rubella among pregnant women in Tanzania. As a result, this study was undertaken to establish the sero-positivity rate of rubella and rubella risk factors among pregnant women attending antenatal care clinics in Mwanza, Tanzania. Methods From November 2012 to May 2013 a total of 350 pregnant women were enrolled and their serum samples collected and analyzed using the AXSYM anti-rubella virus IgG/IgM-MEIA test. Demographic and clinical data were collected using a standardized data collection tool. Data analysis was done using STATA version 12. Results Of 342 pregnant women tested for rubella antibodies, 317 (92.6%) were positive for anti-rubella IgG while only 1 (0.3%) was positive for IgM. Higher sero-positivity rates were found in the age group of 25–44 years. Furthermore, it was observed that with each year increase in age, the risk of contracting rubella increases by 12% (OR = 1.12, 95% CI: 1.02-1.22, P = 0.019). Women involved in farming and business women were at a higher risk of contracting rubella infection compared to formally employed women (OR: 4.9, P = 0.011; OR 7.1, p = 0.003 respectively). In univariate analysis, the risk of contracting rubella virus infection was found to increase with gestational age with a statistical significance. Conclusions Sero-positivity rates of rubella are high in Mwanza and are significantly associated with an increase in age and being a farmer or a business woman. Screening of rubella and immunization of women at risk are highly recommended in this area with a high non-immune rate against rubella virus.