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Reduced false positive reactions in the Dot-enzyme-linked immunosorbent assay for human visceral leishmaniasis
Specificity of the Dot-enzyme-linked immunosorbent assay (Dot-ELISA) for visceral leishmaniasis was significantly improved through the use of enzyme-conjugated antisera specific for IgG heavy chains. Of sera from Kenyans with visceral leishmaniasis, 97% (29/30) were positive using horseradish peroxidase (HRP)-conjugated anti-IgG (heavy and light chain specific) which detected bound IgG and IgM. False positive reactions occurred in 80% of sera from both trypanosomiasis-infected patients (8/10) and apparently healthy Africans (24/30). HRP-conjugated anti-IgG (heavy chain specific), which detected only bound IgG, significantly reduced false positive reactions among trypanosomiasis-infected (2/10, P less than 0.02) and healthy Africans (6/30, P less than 0.001), without reducing test sensitivity in leishmaniasis patients. No false positives occurred when either HRP-conjugated antiserum was used to assay sera from 30 North Americans. Application of enzyme-conjugated antisera specific for IgG improves the serodiagnostic value of the Dot-ELISA for individual patient evaluation and epidemiologic investigations.
Specificity of the Dot-enzyme-linked immunosorbent assay (Dot-ELISA) for visceral leishmaniasis was significantly improved through the use of enzyme-conjugated antisera specific for IgG heavy chains. Of sera from Kenyans with visceral leishmaniasis, 97% (29/30) were positive using horseradish peroxidase (HRP)-conjugated anti-IgG (heavy and light chain specific) which detected bound IgG and IgM. False positive reactions occurred in 80% of sera from both trypanosomiasis-infected patients (8/10) and apparently healthy Africans (24/30). HRP-conjugated anti-IgG (heavy chain specific), which detected only bound IgG, significantly reduced false positive reactions among trypanosomiasis-infected (2/10, P less than 0.02) and healthy Africans (6/30, P less than 0.001), without reducing test sensitivity in leishmaniasis patients. No false positives occurred when either HRP-conjugated antiserum was used to assay sera from 30 North Americans. Application of enzyme-conjugated antisera specific for IgG improves the serodiagnostic value of the Dot-ELISA for individual patient evaluation and epidemiologic investigations.