Enzyme-linked immunosorbent assay and Western blot analysis of dried blood spots (DBS) on filter paper have been shown to be as sensitive and specific as analysis of serum, and therefore may be a cost-effective and culturally appropriate HIV seroprevalence tool in remote areas. This study examines the acceptability of DBS in a tropical, rural population from an outpatient clinic in Andhra Pradesh, India, where participation was offered to every fifth patient seeking general medical care between March and April 2001. All 1413 patients approached for the study agreed to participate and provide a DBS for examination. The overall HIV seroprevalence in this sample was 2.8%. Of the participants, 51.7% were male, 93.2% were between the ages of 18 and 40, 85.3% were married, 29.7% were employed, 47.6% had no education and 73.1% resided in a rural setting. In the univariate analysis, history of genital warts (P = 0.01), sexually transmitted disease (P = 0.001), premarital sexual intercourse (P = 0.002), sexual contact with a commercial sex worker (P = 0.003), being employed (P = 0.011) and having more than 10 injections for medical purposes (P = 0.006) all correlated with being HIV-infected. Given the uniform willingness of these clinic attendees to be tested, we conclude that DBS is a useful, cost-effective tool in HIV serosurveillance in a rural, tropical setting.