The diagnosis of bacterial vaginosis is usually based on clinical criteria including homogeneous vaginal discharge, an elevated vaginal pH, the presence of clue cells, and an amine odor. We have evaluated the vaginal flora and clinical signs for 593 pregnant women. Gardnerella vaginalis, Bacteroides spp., and Mycoplasma hominis were isolated more frequently among women with clinical signs than among those without clinical signs of bacterial vaginosis in multivariable analyses that controlled for other bacteria. To determine the laboratory method that best predicted bacterial vaginosis, we calculated the sensitivity, specificity, and predictive value of positive and negative tests for Gram-stained vaginal smears, gas-liquid chromatography of vaginal fluid, and G. vaginalis cultures compared with clinical signs. G. vaginalis culture was sensitive (92%) and both gas-liquid chromatography (78%) and Gram-stained vaginal smears (62%) were moderately sensitive in identifying women with three of the four clinical signs of bacterial vaginosis. However, the Gram-stained vaginal smear (95%) was more specific than G. vaginalis culture (69%) or gas-liquid chromatography (81%). The predictive value of a positive test was also higher for the Gram staining (76%) than for G. vaginalis culture (41%) or gas-liquid chromatography (48%).