Specimens for chlamydial isolation culture and direct fluorescent antibody cytology (DFA) were collected from 1671 women and children from a trachoma-endemic area in Central Tanzania. Trachoma was graded using the new World Health Organization grading scheme, and 54% of the children and 9% of the women had inflammatory trachoma (TF or TI). DFA, using the presence of five elementary bodies as the criterion for a positive test, had a sensitivity of 88.0% and a specificity of 87.5% compared to culture and a sensitivity of 54.7% and specificity of 92.8% compared to clinical diagnosis. Altogether, 52.9% of those with trachoma grade TF were positive on either or both culture and DFA versus 77.0% of those with TI. Twenty-nine isolates were serotyped; 18 were serovar A, ten were serovar B, and one was serovar Ba. Positive cultures or DFA were obtained in 6.9% of those graded clinically as not having TF or TI and in a smaller number of those without any perceptible evidence of disease. Conversely, organisms could not be demonstrated in a number of people with severe inflammation (TI) even though some became positive after multiple repeated culture. These two findings of infection without disease and disease without evidence of infection suggest the importance of the immunologic response to infection in determining the clinical status. DFA was found to be an appropriate test for future field studies of trachoma. Further studies of those with disease but without agent and of those with agent but without disease will help understand the dynamics of infection and transmission and the role of the immune response in this important blinding disease.