Enzyme-linked immunosorbent assay for shiga toxin and shiga-like toxin II using P1 glycoprotein from hydatid cysts

Abstract

Shiga toxin from Shigella dysenteriae type 1 strains and Shiga-like toxins (SLT)I and II fromEscherichia coli bind to terminal α-D-Galp-(1→4)-D-Galp containing g1ycolipids. Hydatid cyst fluid isolated from sheep infected with Echinococcus granulosus contains a glycoprotein (P1gp) with a terminal α-D-Galp-(1→4)-D-Galp disaccharide. Preparations of Pigp were shown to interact directly with Shiga toxin and to inhibit the binding and cytotoxicity of Shiga toxin to HeLa cells. A sandwich ELISAwas developed using preparations of P1gp as the toxin capture molecule, which, with an appropriate polyclonal antibody, was capable of detecting as little as 80 pg/well Shiga toxin and 132 pg/well SLT-II. Th..