Among 256 patients with acute hepatitis A, 17 (6.6%) had a relapse of the disease between 30 and 90 days after the primary episode. We studied 7 of these patients Serologic testing showed mean alanine aminotransferase levels of 1668 IU/L during the acute stage, 107 IU/L during the early convalescence, and 1027 IU/L during the relapse. Tests for IgM antibody against hepatitis A virus were positive in the 7 patients at the onset of disease, with decreasing levels in 3 of the 4 patients tested during the evolution of the illness. Stools collected during the relapse phase showed hepatitis A virus by immune electron microscopy, radioimmunoassay, and molecular hybridization using a 32P-labeled cDNA-hepatitis A virus probe. Stools collected from 4 of these patients 6 to 12 months after the onset of disease were negative for the virus. The finding of hepatitis A virus in the stool of these patients during the relapse phase strongly implicates hepatitis A virus as the causative agent of the clinical relapse.