HIV infection among women of childbearing age is still increasing in the United States. In most states, HIV testing of women or neonates during pregnancy is not mandatory. The current study assessed HIV prenatal testing practices among obstetrician-gynecologists and primary care physicians listed in a regional physician referral data base in a predominantly rural region. Between December 2000 and March 2001 a 20-question survey was sent by mail to regional physicians in obstetrics/gynecology and primary care regarding physician practice demographics and prenatal HIV testing practices. Of 1116 surveys sent, 431 were returned (38.6% response). Only 42% of physicians offered universal HIV prenatal testing. Factors associated with universal testing (p < 0.5) included obstetrics/gynecology as the practice specialty (90%) physicians' age younger than 50 years, and a practice with predominantly Medicaid or African American patients. Further educational and public health initiatives may be needed to increase nonselective, universal HIV testing in pregnant women.