In situ immunoassay for the assessment of Trypanosoma cruzi interiorization and growth in cultured cells.
Interiorization and multiplication of Trypanosoma cruzi within its host cells are usually assessed by counting parasites in fixed and stained cover slip preparations, a subjective and time-consuming method. Here we describe an immunoenzymatic assay (ELISA) for assessing the number of internalized parasites in infected LLC-MK2 seed on chamber slides (NUNC). ELISA was performed employing a rabbit polyclonal serum against trypomastigote components (MOP) and anti-rabbit IgG conjugated to peroxidase. The bottom of the chamber slide was then detached and processed for quantification of internalized parasites by the conventional method. Data analysis showed a linear relationship between optical densities and number of internalized parasites (r2 = 93.99, p < 0.001). The assay was also efficient to assess inhibition of parasite interiorization induced by the monosaccharide NAc-D-glucosamine.
Piazza
R M
RM
Laboratório de Parasitologia, Instituto Butantan, São Paulo, Brazil.
de Andrade Júnior
H F
HF
Umezawa
E S
ES
Katzin
A M
AM
Stolf
A M
AM
D000818:Animals; D002452:Cell Count; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D011817:Rabbits; D014349:Trypanosoma cruzi / Q000254:growth & development*