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OBJECTIVE: The purpose of this study was to assess the factors associated with acceptance of HIV testing during pregnancy on the part of women receiving prenatal care at public clinics. METHODS: Trained interviewers recruited and interviewed 1,357 women receiving prenatal care at clinics in Florida, Connecticut, and New York City. RESULTS: Eighty-six percent of participants reported having been tested or having signed a consent form to be tested. Acceptance of testing was found to be related to strong beliefs about the benefits of testing, knowledge about vertical transmission, perceived provider endorsement of testing, and social support. Women who declined testing said they did so because they did not perceive themselves to be at risk for HIV (21%) or they faced administrative difficulties (16%) with some aspect of the testing process (for example, scheduling, limited availability of pre-test counselors). CONCLUSIONS: Acceptance rates can be increased when women understand the modes of vertical transmission and the role of medication regimens in preventing transmission; believe that prenatal identification of HIV can promote the health of mother and child; and perceive their providers as strongly endorsing testing. These points can be woven into a brief pre-test counseling message and made a routine component of prenatal care.