Human immunodeficiency virus (HIV), the etiological agent of the acquired immunodeficiency syndrome (AIDS), was treated with either Betadine (povidone-iodine) Solution or Betadine Surgical Scrub. HIV inactivation was analyzed using the viral reverse transcriptase assay or by observing the cytopathic effect produced in HIV-infected, H-9, T-cell cultures. The minimum effective Betadine dose was 0.25% for complete inactivation of HIV that was treated for various time intervals (immediate vortex to ten minutes). The titer of HIV stocks used in these experiments (10(5) TCID50 per mL) was greater than amounts generally detected in clinical specimens. Our results provide a rationale for the use of povidone-iodine as a topical antiseptic against HIV in the clinic or laboratory.