title: Innovations in diagnosis and post-therapeutic monitoring of Chagas disease: Simultaneous flow cytometric detection of IgG1 antibodies anti-live amastigote, anti-live trypomastigote, and anti-fixed epimastigote forms of Trypanosoma cruzi.
abstract: This study developed a remarkable methodological innovation (FC-ATE) which enables simultaneous detection of antibodies specific to the three evolutive forms of Trypanosoma cruzi: live amastigote (AMA), live trypomastigote (TRYPO), and fixed epimastigote (EPI) using a differential fluorescence staining as low (AMA), intermediate (TRYPO), and high (EPI). An outstanding performance (100%) was observed in the discrimination of the chagasic (CH) and non-chagasic (NCH) patients. In the applicability of FC-ATE in the diagnosis of Chagas disease, 100% of the CH samples presented positivity in the percentage of positive fluorescent parasites (PPFP) for all the three forms of T. cruzi. Moreover, 94% of the samples of NCH presented negative values of PPFP with AMA and TRYPO, and 88% with EPI. Samples from the NCH group with false-positive results were those belonging to the leishmaniasis patients. Considering the applicability of this technique in post-therapeutic monitoring of Chagas disease, 100% of non-treated (NT) and treated non-cured (TNC) samples were positive with the three T. cruzi evolutive forms, while a percentage of 100% from samples of the treated cured (TC) patients were negative with AMA, 93% with TRYPO and 96% with EPI. The comparison between FC-ATE and two other flow cytometric tests using the same samples of patients NT, TNC and TC showed that the three techniques presented different reactivities, although categorical correlation between the methodologies was observed. Taken together, the results obtained with the novel FC-ATE method have shown an outstanding performance in the diagnosis and post-therapeutic monitoring of Chagas disease.
authors: Laboratório de Doença de Chagas, Núcleo de Pesquisas em Ciências Biológicas (NUPEB), Instituto de Ciências Exatas e Biológicas (ICEB), Universidade Federal de Ouro Preto (UFOP), Ouro Preto, MG, Brasil.
authors: Glaucia Diniz
authors: GD
authors: Alessio
authors: Laboratório de Doença de Chagas, Núcleo de Pesquisas em Ciências Biológicas (NUPEB), Instituto de Ciências Exatas e Biológicas (ICEB), Universidade Federal de Ouro Preto (UFOP), Ouro Preto, MG, Brasil.
authors: Denise Fonseca
authors: DF
authors: Côrtes
authors: Laboratório de Doença de Chagas, Núcleo de Pesquisas em Ciências Biológicas (NUPEB), Instituto de Ciências Exatas e Biológicas (ICEB), Universidade Federal de Ouro Preto (UFOP), Ouro Preto, MG, Brasil.
authors: Girley Francisco
authors: GF
authors: Machado de Assis
authors: Laboratório de Parasitologia Celular e Molecular, Centro de Pesquisas René Rachou (CPqRR-FIOCRUZ/MG), Belo Horizonte, MG, Brasil.
authors: Policarpo Ademar Sales
authors: PA
authors: Júnior
authors: Universidade Federal de Uberlândia, Instituto de Ciências Biomédicas, Departamento de Morfologia, Uberlândia, MG, Brasil.
authors: Eloisa Amália Vieira
authors: EA
authors: Ferro
authors: Laboratório de Imunopatologia, Centro de Pesquisas René Rachou (CPqRR-FIOCRUZ/MG), Belo Horizonte, MG, Brasil.
authors: Lis Ribeiro do Valle
authors: LR
authors: Antonelli
authors: Laboratório de Biomarcadores de Diagnóstico e Monitoração, Centro de Pesquisas René Rachou (CPqRR-FIOCRUZ/MG), Belo Horizonte, MG, Brasil.
authors: Andréa
authors: A
authors: Teixeira-Carvalho
authors: Laboratório de Biomarcadores de Diagnóstico e Monitoração, Centro de Pesquisas René Rachou (CPqRR-FIOCRUZ/MG), Belo Horizonte, MG, Brasil. Electronic address: oamfilho@cpqrr.fiocruz.br.
authors: Olindo Assis
authors: OA
authors: Martins-Filho
authors: Laboratório de Doença de Chagas, Núcleo de Pesquisas em Ciências Biológicas (NUPEB), Instituto de Ciências Exatas e Biológicas (ICEB), Universidade Federal de Ouro Preto (UFOP), Ouro Preto, MG, Brasil.
authors: Marta
authors: M
authors: de Lana
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000913:Antibodies, Protozoan / Q000097:blood*; D000953:Antigens, Protozoan / Q000097:blood*; D016022:Case-Control Studies; D014355:Chagas Disease / Q000175:diagnosis* / Q000188:drug therapy / Q000276:immunology / Q000469:parasitology; D002648:Child; D002675:Child, Preschool; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood*; D007223:Infant; D008018:Life Cycle Stages / Q000276:immunology*; D008297:Male; D008875:Middle Aged; D011379:Prognosis; D012680:Sensitivity and Specificity; D014344:Trypanocidal Agents / Q000627:therapeutic use; D014349:Trypanosoma cruzi / Q000254:growth & development / Q000276:immunology*
chemical_list: D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D007074:Immunoglobulin G; D014344:Trypanocidal Agents
keywords: Chagas disease; Diagnosis; Flow cytometry; Post-therapeutic monitoring; Serology