Seroepidemiologic and clinical studies of amoebiasis require improved methods for the detection and titration of anti-Entamoeba histolytica (Eh) trophozoite antibodies. We have developed a Dot-enzyme-linked immunosorbent assay (Dot-ELISA) to titrate anti-Eh antibodies in human serum and colostrum. The anti-Eh serum IgG antibody titers from healthy individuals and from patients with amoebic liver abscess obtained, with our Dot-ELISA were similar to those obtained with a standard ELISA, but the background values were lower. Significant anti-Eh sIgA antibody titers were found in seven out of 42 colostrum samples of mothers of newborn children with diarrhea. Our Dot-ELISA is rapid, inexpensive, and might be used for the diagnosis of individuals and in seroepidemiologic studies of amoebiasis.