Navegando por Autor "Santos, Lucas Magno Oliveira"
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Item Immunodiagnosis of human and canine visceral leishmaniasis using recombinant Leishmania infantum Prohibitin protein and a synthetic peptide containing its conformational B-cell epitope.(2019) Rodrigues, Marcella Rezende; Santos, Lucas Magno Oliveira; Miyazaki, Carolina Kei; Martins, Vivian Tamietti; Ribeiro, Fernanda Ludolf; Kursancew, Amanda Christine da Silva; Ramos, Fernanda Fonseca; Dias, Daniel Silva; Oliveira, Jamil Silvano de; Vieira, Paula Melo de Abreu; Roatt, Bruno Mendes; Ávila, Ricardo Andrez Machado de; Gonçalves, Denise Utsch; Souza, Daniel Menezes; Coelho, Eduardo Antônio Ferraz; Duarte, Mariana CostaIn the present study, Leishmania infantum's Prohibitin was cloned and, alongside a synthetic peptide, evaluated for the serodiagnosis of visceral and tegumentary leishmaniasis (CVL and TL, respectively) in dogs and humans. For TL diagnosis, this study analyzed serum samples from cutaneous (n=20) or mucosal (n=39) leishmaniasis patients, and from Chagas disease (CD) patients (n=8) and non-infected patients (n=45). For CVL diagnosis, serum samples from asymptomatic (n=14), symptomatic (n=71), non-infected (n=116), and Leish-Tec®- vaccinated (n=79) dogs were examined, as well as T. cruzi (n=11) and Ehrlichia canis (n=10) infected animals. An indirect ELISA method using rProhibitin showed diagnostic sensitivity and specificity values of 91.76% and 89.91%, respectively. L. infantum SLA showed 86.11% and 48.24% of specificity and sensitivity, respectively, for CVL serodiagnosis, and 98.31% and 84.91% sensitivity and specificity, respectively for TL diagnosis. L. braziliensis SLA showed 75.47% and 83.05% of specificity and sensitivity, respectively, for TL diagnosis. The synthetic peptide showed a better result in TL than in CVL diagnosis. In conclusion, preliminar results suggest that the detection of antibodies against the rProhibitin protein and the synthetic peptide improves the serodiagnosis of TL and CVL.Item Immunoproteomics approach for the discovery of antigens applied to the diagnosis of canine visceral leishmaniasis.(2023) Costa, Scarleth Silva; Santos, Lucas Magno Oliveira; Freire, Larissa Chaves; Tedeschi, Ana Luiza Filizzola; Ribeiro, Naianda Rezende; Queiroz, Mariana Helena Rodrigues; Beraldo Neto, Emídio; Pimenta, Daniel Carvalho; Galvani, Nathália Coral; Luiz, Gabriel Paulino; Oliveira, Maria Eduarda de; Ávila, Ricardo Andrez Machado de; Carvalho, Ana Maria Ravena Severino; Brigido, Bryan Victor Serafim; Reis, Alexandre Barbosa; Fernandes, Ana Paula Salles Moura; Coelho, Eduardo Antônio Ferraz; Roatt, Bruno Mendes; Souza, Daniel Menezes; Duarte, Mariana CostaIn the present study, an immunoproteomic approach using Leishmania infantum parasites isolated from naturally infected dogs from an endemic region of the disease, was carried out to identify new antigens to be used in the diagnosis of canine visceral leishmaniasis (CVL). Protein extracts, obtained from parasites isolated from asymptomatic (CanLA) and symptomatic (CanLS) dogs, were used to perform the two-dimensional gels. Western Blotting assays were carried out by employing a pool of sera from dogs with visceral leishmaniasis (CanLA or CanLS), healthy dogs from an endemic area, or dogs with similar diseases associated with cross-reactions (babesiosis and ehrlichiosis). With these results, it was possible to exclude the spots that showed a crossreactivity of the sera from groups of healthy dogs, and those with babesiosis or ehrlichiosis. Taken together, 20 proteins were identified, 15 of which have already been described in the literature and 5 of which are hypothetical. An immunogenomic screen strategy was applied to identify conserved linear B-cell epitopes in the identified hypothetical proteins. Two peptides were synthesized and tested in ELISA experiments as a proof of concept for the validation of our immunoproteomics findings. The results demonstrated that the antigens presented sensitivity and specificity values ranging from 81.93% to 97.59% and 78.14 to 85.12%, respectively. As a comparative antigen, a preparation of a Leishmania extract showed sensitivity and specificity values of 75.90% and 74.88%, respectively. The present study was able to identify proteins capable of being used for the serodiagnosis of canine visceral leishmaniasis.