BACKGROUND Human immunodeficiency virus (HIV) and syphilis are major public health problems in sub-Saharan Africa, causing numerous adverse pregnancy outcomes. The aim of study was to assess the magnitude of HIV and syphilis seroprevalence among pregnant women at University of Gondar Teaching Hospital. METHOD The study was conducted between March and May, 2012. Sociodemographic data were collected through face-to-face interview. HIV1/2 was tested following current national HIV1/2 testing algorithm. Syphilis infection was also tested using the rapid plasm reagin test for screening and Treponema pallidum hemagglutination as a confirmatory test. Both bivariate and multivariate analysis were used to identify factors associated with HIV and syphilis seroprevalence from selected sociodemographic variables. RESULTS Of 300 women, 31 (10.33%), eleven (3.7%), and three (1%) were seroreactive for HIV, syphilis, and HIV-syphilis coinfection, respectively. High seroprevalence of HIV was found in women ages 25-30 years (13.4%), and women whose husbands attended primary school (19.7%). Syphilis was high in women occupationally housewives (15.2%) and whose husbands were illiterate (11.5%). HIV was associated with husband illiteracy (AOR [adjusted odds ratio] of 4.13, 95% CI [confidence interval] [1.01, 16.95]) and primary educational level of husbands (AOR [95% CI] =3.83 [1.50, 9.90]), whereas syphilis was associated with illiteracy of husband (AOR [95% CI] =7.25 [1.74, 30.30]). CONCLUSION Seroprevalence of HIV and syphilis was high. Low husband educational status was a risk factor for HIV and syphilis. Therefore, substantial efforts have to be made to reinforce prevention strategies and to screen as early as possible to prevent mother-to-child and further horizontal transmission.