title: In situ immunoassay for the assessment of Trypanosoma cruzi interiorization and growth in cultured cells.
abstract: 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.
authors: Laboratório de Parasitologia, Instituto Butantan, São Paulo, Brazil.
authors: R M
authors: RM
authors: Piazza
authors: H F
authors: HF
authors: de Andrade Júnior
authors: E S
authors: ES
authors: Umezawa
authors: A M
authors: AM
authors: Katzin
authors: A M
authors: AM
authors: Stolf
mesh_terms: D000818:Animals; D002452:Cell Count; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D011817:Rabbits; D014349:Trypanosoma cruzi / Q000254:growth & development*