Shigella enterotoxin 1 (ShET1) is a novel, iron-dependent, toxin encoded by chromosomal genes (set1). To determine the prevalence of this enterotoxin, 172 Shigella clinical isolates (and 10 enteroinvasive Escherichia coli [EIEC]) from distant areas worldwide, representing all 4 groups and 45 serotypes of Shigella, were screened for set1 by DNA colony hybridization and polymerase chain reaction amplification. set1 was present in all 22 Shigella flexneri 2a strains tested but was rare in isolates of other Shigella serotypes (3.3%, 5/150) and not found in EIEC (0/10). That ShET1 is found almost exclusively in S. flexneri 2a may help explain the epidemiologic predominance of this serotype in the developing world.