Serological diagnosis of Chagas disease: evaluation and characterisation of a low cost antigen with high sensitivity and specificity

Campos, Yelitza; Briceño, Luis; Reina, Kate; Figarella, Katherine; Pérez, José Luís; Mosca, W

doi:10.1590/s0074-02762009000600016

Cited by 10 publications

(12 citation statements)

References 7 publications

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“…12,[17][18][19][20][21][22][23] The main limitation of this technique is that it is difficult to perform interlaboratory comparisons, mainly due to the heterogeneity of the strains and antigenic preparations used. In addition, the size of the slab gels and the concentration of polyacrylamide used as well as the different molecular weight markers all introduce additional variations in band separation and the establishment of respective molecular weights.…”

Section: Discussionmentioning

confidence: 99%

Identification of a Western Blot Pattern for the Specific Diagnosis of Trypanosoma cruzi Infection in Human Sera

Riera

Vergés²,

Iniesta³

et al. 2012

The American Society of Tropical Medicine and Hygiene

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Abstract. A Western blot (WB) method using a lysate from Trypanosoma cruzi (Maracay strain) epimastigotes was evaluated. Serum samples from 37 patients with confirmed Chagas disease (cohort I), 27 Spanish patients with visceral leishmaniasis caused by Leishmania infantum (cohort II), and 28 Colombian patients with cutaneous leishmaniasis caused by L. panamensis and negative serology for Chagas disease (cohort III) were tested. The negative controls were 55 healthy seronegative subjects for T. cruzi and Leishmania; 28 of the negative controls were from a region endemic for Chagas disease and Leishmania (cohort IV), and 27 of the negative controls were from a non-endemic area for Leishmania and T. cruzi (cohort V). A homogeneous standard band pattern consisting of six antigenic bands corresponding to 28, 32, 38, 39, 40, and 48 kDa was recognized simultaneously for all Chagasic patients' sera. Sera from Leishmania-infected patients showed a heterogeneous band pattern that was easily differentiated from the pattern of patients with Chagas disease. WB with T. cruzi epimastigote antigen is an efficient method for diagnosis and may be used as an alternative to confirm T. cruzi and detect cross-reactivity with Leishmania.

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Section: Discussionmentioning

confidence: 99%

Identification of a Western Blot Pattern for the Specific Diagnosis of Trypanosoma cruzi Infection in Human Sera

Riera

Vergés²,

Iniesta³

et al. 2012

The American Society of Tropical Medicine and Hygiene

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“…Furthermore, reactivity is retained for approximately one year after production. Another advantage of ESEA antigens compared to other antigenic preparations from epimastigotes is the easy production since they donot need to be sonicated or follow other biochemical procedures to obtain usable antigenic preparations [20, 30, 31]. …”

Section: Discussionmentioning

confidence: 99%

Development and Application of an ELISA Assay Using Excretion/Secretion Proteins from Epimastigote Forms ofT. cruzi(ESEA Antigens) for the Diagnosis of Chagas Disease

Berrizbeitia

Figueroa

Ward

et al. 2012

Journal of Tropical Medicine

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An indirect enzyme-linked immunoabsorbent assay (ELISA) for Trypanosoma cruzi was developed using epimastigote secretion/excretion proteins (ESEA antigens) obtained from axenic culture supernatants. A panel of 120 serum samples from subjects with confirmed Chagas disease (n = 50), healthy controls (n = 50), and patients with other parasitic diseases (n = 20) was used to evaluate the new ESEA-based ELISA (ELISAESEA). This new test had excellent sensitivity (98%) and acceptable specificity (88%). Cross-reactivity was observed largely in sera from subjects with Leishmania and Ascaris infections. Using Western blotting and epimastigotes from two distinct T. cruzi isolates, several polypeptide bands with molecular masses ranging from 50 to 220 kDa were detected in pooled chagasic sera. However, the band pattern for each isolate was different. These data suggest that an inexpensive and technically simple ELISA based on ESEA antigens is a promising new tool for the diagnosis of Chagas disease.

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“…T. cruzi parasite culture - Epimastigote forms of the CL Brener T. cruzi strain were cultured in vitro at 27ºC using Schenider’s Insect Medium at pH 6.9, sterilised by filtration and supplemented with 10% fetal bovine serum (Campos et al 2009). Approximately, 1x10 7 parasites (counted in a Neubauer chamber) were collected in logarithmic growth phase.…”

Section: Methodsmentioning

confidence: 99%

Nicotinamide mononucleotide adenylyltransferase of Trypanosoma cruzi (TcNMNAT): a cytosol protein target for serine kinases

Sánchez-Lancheros

Ospina-Giraldo

Ramírez-Hernández

2016

Mem. Inst. Oswaldo Cruz

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Nicotinamide/nicotinate adenine dinucleotide (NAD+/NaAD) performs essential functions in cell metabolism and energy production due to its redox properties. The nicotinamide/nicotinate mononucleotide adenylyltransferase (NMNAT, EC 2.7.7.1/18) enzyme catalyses the key step in the biosynthesis of NAD+. Previously, the enzyme NMNAT was identified in Trypanosoma cruzi (TcNMNAT), a pathogenic agent with epidemiological importance in Latin America. To continue with the functional characterisation of this enzyme, its subcellular location and its possible post-translational modifications were examined in this study. For this, polyclonal antibodies were generated in mice, with soluble and denatured recombinant protein being used to detect the parasite’s NMNAT. Immunodetection assays were performed on whole extracts of T. cruzi, and an approximation of its intracellular location was determined using confocal microscopy on wild and transgenic parasites, which revealed the cytosol distribution patterns. This localisation occurs according to the needs of the dinucleotides that exist in this compartment. Additionally, a bioinformatics study was performed as a first approach to establish the post-translational modifications of the enzyme. Possible phosphorylation events were experimentally analysed by western blot, highlighting TcNMNAT as a potential target for serine kinases.

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Serological diagnosis of Chagas disease: evaluation and characterisation of a low cost antigen with high sensitivity and specificity

Cited by 10 publications

References 7 publications

Identification of a Western Blot Pattern for the Specific Diagnosis of Trypanosoma cruzi Infection in Human Sera

Identification of a Western Blot Pattern for the Specific Diagnosis of Trypanosoma cruzi Infection in Human Sera

Development and Application of an ELISA Assay Using Excretion/Secretion Proteins from Epimastigote Forms ofT. cruzi(ESEA Antigens) for the Diagnosis of Chagas Disease

Nicotinamide mononucleotide adenylyltransferase of Trypanosoma cruzi (TcNMNAT): a cytosol protein target for serine kinases

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