Chagatest ELISA lisado is an "in vitro" enzyme immunoassay for the qualitative detection of antibodies anti-T. cruzi in
human serum or plasma samples.
The sample is diluted in the wells in which T.cruzi antigens are
immobilized, corresponding to highly preserved zones among
different strains. If the sample contains specific antibodies,
they will bind to the antigens and remain bound to the solid
phase. Unbound fractions are eliminated by washing, after
which conjugate is added (monoclonal antibodies anti-human
IgG, conjugated to peroxidase), which specifically reacts
with immunocaptured antibodies anti-T.cruzi. The unbound
conjugate is removed by washing. The presence of peroxidase bound to the complex is recognized by the addition of
a chromogenic substrate, tetramethylbenzidine. The reactive
samples develop a light blue color. The enzymatic reaction
is stopped by the addition of sulphuric acid, producing a light
blue to yellow color change. The optical density is measured
bichromatically at 450/620-650 nm or at 450 nm.
Coated microtitration plate: microtitration plate with removable strips and 96 wells containing Trypanosoma cruzi
Sample Diluent: saline buffer with surfactant. Violet color.
Concentrated Conjugate: monoclonal antibody anti-Human
IgG, conjugated to peroxidase (10x). Red color.
Conjugate Diluent: saline buffer with proteins.
Substrate: tetramethylbenzidine and hydrogen peroxide solution.
Stopper: 2 N sulfuric acid.
Concentrated Wash Buffer: saline buffer with surfactant
(25x). Green color.