Improved enzyme-linked immunoadsorbent assay (ELISA) for the study of Trypanosoma cruzi-host cell interaction in vitro.
We herein present an improved assay for detecting the presence of Trypanosoma cruzi in infected cultures. Using chagasic human sera (CHS), we were able to detect T. cruzi infection in primary cultures of both peritoneal macrophages and heart muscle cells (MHC). To avoid elevated background levels--hitherto observed in all experiments especially in those using HMC--CHS were preincubated with uninfected cells in monolayers or suspensions prior to being used for detection of T. cruzi in infected monolayers. Preincubation with cell suspensions gave better results than with monolayers, reducing background by up to three times and increasing sensitivity by to twenty times. In addition, the continuous fibroblastic cell line L929 was shown to be suitable for preadsorption of CHS. These results indicate that the high background levels observed in previous reports may be due to the presence of human autoantibodies that recognize surface and/or extracellular matrix components in cell monolayers. We therefore propose a modified procedure that increases the performance of the ELISA method, making it an useful tool even in cultures that would otherwise be expected to present low levels of infection or high levels of background.
Luz
M R
MR
Departamento de Ultraestrutura e Biologia Celular, Instituto Oswaldo Cruz, Rio de Janeiro, Brasil.
Soeiro
M de N
Mde N
Araujo-Jorge
T C
TC
D000818:Animals; D002448:Cell Adhesion; D014355:Chagas Disease / Q000175:diagnosis*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006790:Host-Parasite Interactions; D066298:In Vitro Techniques; D008264:Macrophages / Q000469:parasitology*; D008297:Male; D051379:Mice; D010529:Peritoneal Cavity / Q000166:cytology; D014349:Trypanosoma cruzi / Q000502:physiology*