Double-antibody sandwich ELISA using biotinylated antibodies for the detection of Echinococcus granulosus coproantigens in dogs

Abstract

Here we present the diagnostic evaluation of an improved double-antibody sandwich ELISA for detecting Echinococcus granulosus antigens in dog faecal samples (coproantigens). A purified rabbit IgG fraction against protoscolex excretory-secretory products was used as primary antibody, and the same fraction conjugated with biotin as secondary antibody. In order to validate the sandwich ELISA, intra- and inter-assay precision, linearity, and recovery percentages were calculated. The diagnostic evaluation of the method was carried out by investigating faecal samples from 37 dogs naturally infected with E. granulosus, 15 Echinococcus-free dogs infected with Taenia spp., 82 dogs with non-taeniid helminths and 66 dogs free of helminth infections. An overall sensitivity of 78.4% and specificity of 93.3% were determined. Positive and negative predictive values were 72 and 95%, respectively, and the diagnostic efficiency was 90.5%. In addition, the sandwich ELISA detection limit was estimated in 5.12 ng ml(-1). These results are highly satisfactory, allowing the use of this methodology in surveillance and control programs for intestinal echinococcosis in dogs.