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Infection with syphilis during pregnancy could cause spontaneous abortion, low birth weight and stillbirth. To prevent adverse pregnancy outcomes caused by syphilis, the World Health Organization (WHO) recommends syphilis screening and treatment of all pregnant women attending antenatal care (ANC) services. Rapid syphilis testing (RST) demonstration project was introduced at ANC clinics in Tanzania in 2009, to assess the feasibility, acceptability and its impact on uptake of syphilis screening service. Data collection was composed of in-depth interviews with health workers and pregnant women attending ANC. Additionally, from the health facility registers we extracted information on the uptake of antenatal care services, including number of pregnant women screened and treated for syphilis. Introduction of RST at health facilities was appreciated by pregnant women attending ANC and health workers. Following the introduction of RST services at ANC clinics, we observed a significant increase of the uptake of syphilis screening. Pregnant women appreciated RST service since it reduced the frequency of their visits to the health facilities and shortened the duration that they spent at the clinics. Moreover, the provision of same-day screening and treatment services helped women to save money that they would have to spend on transportation for the follow up visits. Health workers felt that RST simplified procedures to diagnose syphilis, and enabled the health workers to test and treat large numbers of clients in a shorter period of time. Our study demonstrates that, it is feasible to introduce RST service in antenatal clinics. The RST was appreciated by health workers and pregnant women, since it simplifies syphilis screening procedures, saves the time that pregnant women used to waste to wait for the results, and saves the cost that women would have to spend on transportation to come back for treatment.