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. 2018 Sep 11;40(11):e12584. doi: 10.1111/pim.12584

Figure 3.

Figure 3

Antigen detection capture ELISA assembled with VHHs for the identification of Li‐isd1 in urine of VL patients and controls. ELISA plates were coated with either VHH JRD‐C1/JRD‐C1‐C1/myc (2 μg/mL) or antigen affinity‐purified IgY anti‐Li‐isd1 antibody (1 μg/mL). After overnight incubation with patients or control, urine samples plates were washed and wells were incubated with E‐tag‐labelled developing VHH JRD‐E5/JRD‐E9 or biotin‐labelled rabbit IgG anti‐Li‐isd1 antibody. Wells corresponding to reactions with VHHs were next incubated with peroxidase‐labelled rabbit anti‐E‐tag antibody. Wells incubated with biotinylated IgG were next incubated with streptavidin‐peroxidase. The substrate H2O2 and the chromophore TMB were then added followed by OD reading at 450 nm. Samples from VL patients and from controls were from Belo Horizonte, MG, Brazil. Dashed lines represent the cut‐off values calculated as described in the text. These are representative results of at least three experiments performed at different times with the same urine samples and same capture ELISA. VL, visceral leishmaniasis