Abstract: Objective To study the awareness of HIV/AIDS and syphilis related knowledge among early pregnant women and to understand its influencing factors on accepting screening tests. Methods Early pregnant women, who were residents in Yuexiu District of Guangzhou, were surveyed with questionnaire from February 2012 to January 2013. The contents of questionnaire included demographic characteristics，awareness of HIV/AIDS and syphilis transmission related knowledge，prevention, attitude and behavior on screening test for pregnant women．Chi-square test and multivariate logistic regression were used for data analysis. Results A total of 6 012 pregnant women were surveyed. Their average age was（29.07±3.57）years, 60.83% of them were bachelor degree, and 38.31% of their jobs were cadre or staff. Of them, 81.74% understood general knowledge of HIV/AIDS and syphilis, 76.72% understood the transmission route of AIDS and syphilis, 78.69% knew the mother-to-child transmission（MTCT）, and awareness rate of preventive measures was 80.82%. Although 81.90% of the respondents expressed their willingness to participate in the free screening tests of HIV and syphilis antibodies, only 55.19% of them had their blood tested for free testing on anti-HIV and treponema pallidum particle agglutination (TPPA). Multivariate regression showed that pregnant women who did not have premarital medical check-up had more possibility to accept the related testing compared with those who did (OR =1.207) , pregnant women who had never detected HIV antibodies had higher possibility to accept HIV antibodies detected than those who previously had HIV antibodies detected (OR =1.805), and the possibility of accepting free testing in those who expressed unwilling to accept the free test was 0.548 times compared with those who expressed willing to do it. Conclusion The awareness rate of HIV and syphilis MTCT knowledge of pregnant women is relatively low. Health education should be strengthened in early pregnant women to improve their awareness level of MTCT and preventive measures. Multi-level testing service system should be established to promote test rate during the first and second trimesters of pregnancy to prevent MTCT.