Fig 1.
Groups of patients included in this study.
It was defined the cases or non-cases of American Tegumentary Leishmaniasis (ATL) from suspected patients by four diagnostic methods mentioned above. Three of those 129 subjects were inconclusive and could not be grouped. Additionally, for the non-ATL cases, there were included two groups of donors without ATL suspicion: Chagas seropositives and Japanese donors from non-endemic geographic area for ATL and Chagas disease.
Table 1.
Leishmania zymodemes used for the preparation of antigenic crude extracts.
Fig 2.
Comparison between the main local species, promastigoste vs. amastigote extracts and membrane based vs. soluble extracts.
Comparison between Optical Density (OD) for each ATL case and resultant absorbance means (ODm) obtained from the following three consecutive assays, showing the differences among the avidity for anti-Leishmania antibodies. A) The ODs of 17 ATL cases and its ODm obtained by ELISA using promastigote Ags of five Leishmania zymodemes (0.15 μg/well), sera (1/2000,100 μl/well), and conjugate (1/500) conditions [*p by ANOVA/Dunn´s multiple comparison test vs. main local L. (V.) braz]; B) The ODs of 21 ATL cases obtained by using amastigote or promastigote Ags from Argentinean L. (V.) braziliensis main zymodeme (0.15 μg/well), sera (1/2000,100 μl/well), and conjugate (1/500) [*p by Wilcoxon test]; C) The ODs of 21 ATL cases obtained by using membrane or soluble extracts from amastigotes of Argentinean L. (V.) braziliensis main zymodeme under conditions previously described [*p by Wilcoxon test]. A p-value less than 0.05 was considered significant.
Fig 3.
Diagnostic performance of the selected antigenic extract.
Enzyme Lynked Immunosorbent Assay reactivity using the membrane amastigote extract against ATL and non-ATL cases sera Aand B, and excluding Chagatest reactive samples, C and D. The relation / CO, where
is the average of duplicate or triplicate ODs for each sample tested, and CO = Cut-off. The indetermination area (IA) is shown in dotted line, and defined as CO / CO±10%CO / CO = 0,9–1,1. B and D show the Receiver Operating Characteristic (ROC) Curves for ELISAs.
Table 2.
The performance of ELISA using amastigote-membrane extracts of ‘local’ L. (V.) braziliensis as antigens.