Characterization of monoclonal antibodies to conserved antigens of Entamoeba histolytica and E. dispar and development of a stool ELISA

Abstract

Eight murine monoclonal antibodies (MAbs) were generated against Entamoeba histolytica NIH:200. All MAbs reacted with three axenic strains of E. histolytica, NIH:200, HM-1: IMMS, and SAW 1734R clAR, but not with enteric bacteria or Giardia lamblia. Five of the MAbs reacted with low-molecular-weight, periodate-sensitive antigens of 14-21 kD, while one (AB31) reacted with high-molecular-weight, 90 to 200-kD protein determinants. MAb PC14 appeared to be specific for antigen in its native state. Another MAb (BB12) agglutinated live trophozoites and caused membrane fluorescence in contrast to the five other MAbs tested. Although BB12 reacted with the same 14 to 21-kD band on Western blot as AC55, the latter reacted with different cytoplasmic epitopes. All the MAbs reacted to five pathogenic (E. histolytica) and six nonpathogenic (E. dispar) clinical isolates. These MAbs may be helpful for studying conserved antigens of E. histolytica and were used to develop a sandwich ELISA for the diagnosis of intestinal amoebiasis. The assay was sensitive to 60 ng of E. histolytica antigen. A comparative study of microscopic examination of stool samples and the sandwich ELISA was conducted on 102 samples from patients with gastrointestinal complaints. The ELISA could detect all microscopically positive samples with a sensitivity of 100% and specificity of 93%. A sandwich ELISA using a monoclonal antibody to conserved antigens of E. histolytica has the potential to be a reliable method for the diagnosis of intestinal amoebiasis.