Antiretroviral therapy during pregnancy in HIV-infected women has dramatically reduced the rate of mother to child HIV transmission in the United States. National guidelines strongly recommend universal HIV testing of all pregnant women with repeat screening in the third-trimester in high-risk populations. To determine patient attitudes towards third-trimester rescreening, a convenience sample was recruited during routine prenatal visits at an urban clinic and participants were surveyed to determine attitudes about HIV third-trimester retesting, acceptability of the rapid HIV testing, condom use, and knowledge of partner's HIV status during pregnancy. Participants were offered a third-trimester rapid HIV retest with the option to decline the test. Eighty pregnant women participated; 95% agreed to be retested with a rapid HIV test, 100% received immediate HIV results, and 91% reported that the rapid test was less stressful than conventional testing. There were no seroconversions. Although 35% did not know their partner's HIV status, 57% of these women reported never using condoms during pregnancy. There was a significant association between reported stage of behavior change and reported likelihood of using condoms. We found that rescreening with the rapid HIV test in the third trimester of pregnancy was well accepted and is important to prevent perinatal HIV transmission.