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Pregnant women infected with HCV should be given attention due to their special physiological stage and the effect on offspring health. To examine the prevalence of HCV infection among pregnant women in part of China and explore relevant factors during pregnancy, a cross-sectional study was conducted in four maternal and children health care institutions (MCHC) in Guangdong, Hunan and Chongqing. Pregnant women who were delivered, induced or spontaneous abortion were included and relevant information was collected through the Hospital Information System. Results showed that the prevalence of HCV among pregnant women in four MCHCs was 0.11% (95% CI 0.09–0.13%). Age, occupations, regions, syphilis-infection, intrahepatic cholestasis of pregnancy (ICP), and placenta previa were significant factors (all P < 0.05). Age and syphilis-infection were positively correlated with HCV infection (Z = 3.41, P = 0.0006; OR = 18.16, 95% CI 9.34–35.29). HCV and HBV infection were risk factors of ICP (OR = 4.18, 95% CI 2.18–8.04; OR = 2.59, 95% CI 2.31–2.89). Our study indicates that the prevalence of HCV among pregnant women in the three provinces(city) was low compared with the general population in China. Older age and syphilis-infection increased the risk of HCV infection during pregnancy. HCV infection was a risk factor of ICP. Generally, we need keep a watchful eye on HCV infection and relevant factors mentioned above during pregnancy in clinic, especially those also infected with syphilis. HCV testing based on risk factors is recommended in antenatal care and obstetrics. [ABSTRACT FROM AUTHOR]