Immunoblotting analyses using two-dimensional gel electrophoresis of Trypanosoma cruzi excreted-secreted antigens

Revista da Sociedade Brasileira de Medicina Tropical

Adriano Gomes Silva²  Elisangela Paula Silveira-lacerda¹  Jair Pereira Cunha-júnior²  Maria Aparecida De Souza²  Silvio Favoreto Junior² 

[1] ,Universidade Federal de Uberlândia Instituto de Ciências Biomédicas Laboratório de ImunologiaUberlândia MG

关键词: Trypanosoma cruzi;    Excreted-secreted antigens;    Two-dimensional gel electrophoresis;    Immunoblotting;    Trypanosoma cruzi;    Antígenos excretados;    Eletroforese em duas dimensões;    Immunoblotting;   

DOI  :  10.1590/S0037-86822004000600005

来源: SciELO

【 摘 要 】

Trypanosoma cruzi trypomastigotes excrete-secrete a complex mixture of antigenic molecules. This antigenic mixture denominated trypomastigote excreted-secreted antigens contains a 150-160 kDa band that shows excellent performance in Chagas' disease diagnosis by immunoblotting. The present study partially characterized by two-dimensional gel electrophoresis the immunoreactivity against the 150-160kDa protein using sera samples from chagasic patients in different phases of the disease. Trypomastigote excreted-secreted antigen preparations were subjected to high-resolution two-dimensional (2D) gel electrophoresis followed by immunoblotting with sera from chagasic and non-chagasic patients. The 150-160kDa protein presented four isoforms with isoelectric focusing ranging from 6.2 to 6.7. The four isoforms were recognized by IgM from acute phase and IgG from chronic phase sera of chagasic patients. The 150-160kDa isoform with IF of approximately 6.4 became the immunodominant spot with the progression of the disease. No cross-reactivity was observed with non-chagasic or patients infected with Leishmania sp. In this study we provide basic knowledge that supports the validation of trypomastigote excreted-secreted antigens for serological diagnosis of Chagas' disease.

【 授权许可】

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