ABSTRACT
Protein tyrosine phosphatases (PTPs) play an essential role in the regulation of cell differentiation in pathogenic trypanosomatids. In this study, we describe a PTP expressed by the non-pathogenic protozoan Trypanosoma rangeli (TrPTP2). The gene for this PTP is orthologous to the T. brucei TbPTP1 and Trypanosoma cruzi (TcPTP2) genes. Cloning and expression of the TrPTP2 and TcPTP2 proteins allowed anti-PTP2 monoclonal antibodies to be generated in BALB/c mice. When expressed by T. rangeli epimastigotes and trypomastigotes, native TrPTP2 is detected as a ~65 kDa protein associated with the parasite's flagellum. Given that the flagellum is an important structure for cell differentiation in trypanosomatids, the presence of a protein responsible for tyrosine dephosphorylation in the T. rangeli flagellum could represent an interesting mechanism of regulation in this structure.
Subject(s)
Animals , Mice , Antibodies, Monoclonal/immunology , Flagella/enzymology , Protein Tyrosine Phosphatases/metabolism , Trypanosoma rangeli/enzymology , Immunization , Mice, Inbred BALB C , Phylogeny , Protein Tyrosine Phosphatases/genetics , Trypanosoma rangeli/genetics , Trypanosoma rangeli/immunologyABSTRACT
An indirect enzyme-linked immunosorbent assay (ELISA) using a crude antigen was evaluated for its performance to detect Babesia bigemina antibodies. The sensitivity and specificity were 98.0 per cent and 99.0 per cent, respectively. In agreement with the high specificity, no cross-reactions were verified with sera from calves inoculated three times with 10 7(subscribe) Babesia bovis organisms. With regard to the comparison of ELISA and indirect fluorescent antibody test (IFAT) in detecting antibodies against B. bigemina in calves experimentally infected with five Brazilian geographical isolates of this hemoparasite, IFAT was able to detect antibodies one day earlier in most of the calves' sera. There was a good agreement between results shown by ELISA and IFAT with sera from an enzootically stable area (k=0.61). However, there was no agreement between these serological tests with sera from an enzootically unstable area (k=0.33). The ELISA was employed in an epidemiological survey using with 1,367 sera from four counties in the Pantanal of Mato Grosso do Sul and characterized this region as an enzootically stable area, since the prevalence ranged from 87.7 to 98.9 per cent. Therefore, this ELISA with high sensitivity, specificity and performance similar to IFAT can be employed in serological diagnosis of B. bigemina