AVALIAÇÃO DO DESEMPENHO DIAGNÓSTICO DE NOVAS TÉCNICAS SOROLÓGICAS PARA LEISHMANIOSE VISCERAL CANINA

Detalhes bibliográficos
Autor(a) principal: NATHALIA LOPES FONTOURA MATEUS
Data de Publicação: 2022
Tipo de documento: Tese
Idioma: por
Título da fonte: Repositório Institucional da UFMS
Texto Completo: https://repositorio.ufms.br/handle/123456789/5389
Resumo: Visceral leishmaniasis (VL) is a zoonosis caused by the protozoan Leishmania infantum. Dogs are considered important urban reservoir for the parasite and canine visceral leishmaniasis (CVL) diagnosis contributes for monitoring and reemergence of VL. CVL confirmatory diagnosis as recommended by the Mistry of Health is performed using enzyme-linked immunosorbent assay (ELISA) with crude antigen of L. major-like. However, this technique may provide false positive results for animals infected with other trypanosomatids or Ehrlichia canis. Besides that, false negative results may be obtained for asymptomatic dogs, which can act as the parasite reservoir in the urban environment. In this context, recombinant antigens have been developed individually or combined to increase sensitivity, specificity, and accuracy of CVL serodiagnosis. This study analyzed three serologic tests for CVL diagnosis: two national developed ELISA using recombinant proteins (Q1 and Q5) and one rapid test using antigen of L. donovani promastigote membrane, in dipstick presentation. To evaluate chimeric proteins performances in ELISA, samples from a serological bank were used. Samples from naturally infected dogs with L. infantum symptomatic (n=30), oligosymptomatic (n=30) and asymptomatic (n=10); healthy (n=30), vaccinated with Leish-Tec (n=5) and Leishmune  (n=5) and infected with E. canis (n=30) were used. The performance of chimeric ELISAs were compared to the ELISA with L. infantum total extract as antigen. Greater values of sensitivity and specificity were observed, respectively, for Q1 (81.43%; 85.71%) and Q5 (74.29%; 90.0%), when compared to total extract antigen (71.43%; 82.86%). Although Q1 presented cross-reaction with Leish-Tec-vaccinated dogs, this chimeric protein demonstrated better sensitivity to diagnose asymptomatic (70%) and oligosymptomatic (73,33%) dogs. To access dipstick performance, samples from naturally infected dogs with L. infantum symptomatic (n=15), oligosymptomatic (n=15) and asymptomatic (n=9); healthy (n=15); vaccinated with Leish-Tec (n=5) and infected with E. canis (n=15) were obtained from a sera bank. The dipstick test presented higher sensitivity in the oligosymptomatic group (93.33%); followed by asymptomatic (77.88%) and symptomatic (73.33%); specificity value of 100% was observed for the test among all groups. Therefore, Q5 may be an interesting antigen for the development of serological tests and dipstick test could be a useful tool for CVL diagnosis due to its ease of performance. Both methods demonstrated good performance in situations where canine ehrlichiosis may be a differential diagnosis for the patient.
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spelling 2022-11-30T19:18:41Z2022-11-30T19:18:41Z2022https://repositorio.ufms.br/handle/123456789/5389Visceral leishmaniasis (VL) is a zoonosis caused by the protozoan Leishmania infantum. Dogs are considered important urban reservoir for the parasite and canine visceral leishmaniasis (CVL) diagnosis contributes for monitoring and reemergence of VL. CVL confirmatory diagnosis as recommended by the Mistry of Health is performed using enzyme-linked immunosorbent assay (ELISA) with crude antigen of L. major-like. However, this technique may provide false positive results for animals infected with other trypanosomatids or Ehrlichia canis. Besides that, false negative results may be obtained for asymptomatic dogs, which can act as the parasite reservoir in the urban environment. In this context, recombinant antigens have been developed individually or combined to increase sensitivity, specificity, and accuracy of CVL serodiagnosis. This study analyzed three serologic tests for CVL diagnosis: two national developed ELISA using recombinant proteins (Q1 and Q5) and one rapid test using antigen of L. donovani promastigote membrane, in dipstick presentation. To evaluate chimeric proteins performances in ELISA, samples from a serological bank were used. Samples from naturally infected dogs with L. infantum symptomatic (n=30), oligosymptomatic (n=30) and asymptomatic (n=10); healthy (n=30), vaccinated with Leish-Tec (n=5) and Leishmune  (n=5) and infected with E. canis (n=30) were used. The performance of chimeric ELISAs were compared to the ELISA with L. infantum total extract as antigen. Greater values of sensitivity and specificity were observed, respectively, for Q1 (81.43%; 85.71%) and Q5 (74.29%; 90.0%), when compared to total extract antigen (71.43%; 82.86%). Although Q1 presented cross-reaction with Leish-Tec-vaccinated dogs, this chimeric protein demonstrated better sensitivity to diagnose asymptomatic (70%) and oligosymptomatic (73,33%) dogs. To access dipstick performance, samples from naturally infected dogs with L. infantum symptomatic (n=15), oligosymptomatic (n=15) and asymptomatic (n=9); healthy (n=15); vaccinated with Leish-Tec (n=5) and infected with E. canis (n=15) were obtained from a sera bank. The dipstick test presented higher sensitivity in the oligosymptomatic group (93.33%); followed by asymptomatic (77.88%) and symptomatic (73.33%); specificity value of 100% was observed for the test among all groups. Therefore, Q5 may be an interesting antigen for the development of serological tests and dipstick test could be a useful tool for CVL diagnosis due to its ease of performance. Both methods demonstrated good performance in situations where canine ehrlichiosis may be a differential diagnosis for the patient.A leishmaniose visceral (LV) é uma zoonose causada pelo protozoário Leishmania infantum. Os cães são considerados importantes reservatórios urbanos do parasita e o diagnóstico da leishmaniose visceral canina (LVC) contribui no monitoramento de transmissão e emergência de LV. A confirmação da LVC instituída pelo Ministério da Saúde consiste na reação de imunoadsorção enzimática (ELISA), por meio da utilização de antígeno bruto de L. major-like; porém, esta técnica pode proporcionar resultados falsos positivos em animais infectados por outros tripanossomatídeos ou por Ehrlichia canis. Além disso, resultados falsos negativos são relatados em animais assintomáticos, promovendo a manutenção de reservatórios no ambiente. Neste âmbito, foram desenvolvidos antígenos recombinantes usados de maneira individual ou combinada no diagnóstico sorológico da LVC com intuito de aumentar a sensibilidade, especificidade e acurácia do teste. Este trabalhou avaliou três testes sorológicos para o diagnóstico de LVC: dois ELISA com proteínas recombinantes quiméricas (Q1 e Q5) de desenvolvimento nacional e um teste rápido utilizando antígeno de membrana de promastigota de Leishmania donovani na apresentação de fitas reagentes (dipstick). Para avaliação das proteínas quiméricas, utilizou-se amostras de um banco sorológico provenientes de cães naturalmente infectados por L. infantum sintomáticos (n=30), oligossintomáticos (n=30), assintomáticos (n=10), cães saudáveis (n=30), vacinados com Leish-Tec (n=5) e Leishmune  (n=5) e infectados com Ehrlichia canis (n=30). O resultado dos ELISA utilizando quimeras foi comparado ao ELISA com antígeno de extrato total de L. infantum e observou-se valores maiores de sensibilidade e especificidade, respectivamente, para Q1 (81,43%; 85,71%) e Q5 (74,29%; 90,0%), quando comparado ao antígeno bruto (71.43%; 82.86%). Apesar da proteína Q1 ter apresentado reação cruzada com amostras de cães vacinados com Leish-Tec, esta quimera apresentou melhor sensibilidade no diagnóstico de animais assintomáticos (70%) e oligossintomáticos (73,33%).Para avaliação do teste rápido dipstick foram selecionadas amostras de um banco sorológico provenientes de cães naturalmente infectados por L. infantum sintomáticos (n=15), oligossintomáticos (n=15), assintomáticos (n=9), saudáveis (n=15), vacinados com Leish-Tec (n=5) e infectados com Ehrlichia canis (n=15). O teste apresentou maior sensibilidade no grupo oligossintomático (93,33%), seguido do assintomático (77,78%) e sintomático (73,33%) e foi observado 100% de especificidade em todos os grupos. Desta maneira, Q5 pode ser um antígeno interessante para o desenvolvimento de testes sorológicos e as fitas reagentes (dipstick) podem ser uma ferramenta útil no diagnóstico da LVC devido a praticidade de realização. Ambos os métodos demonstraram bons resultados em situações em que a erliquiose canina pode ser diagnóstico diferencial do paciente.Fundação Universidade Federal de Mato Grosso do SulUFMSBrasilELISA, fitas reagentes, proteína recombinanteproteína quimera, teste rápidoAVALIAÇÃO DO DESEMPENHO DIAGNÓSTICO DE NOVAS TÉCNICAS SOROLÓGICAS PARA LEISHMANIOSE VISCERAL CANINAinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisCarlos Alberto do Nascimento RamosNATHALIA LOPES FONTOURA MATEUSinfo:eu-repo/semantics/openAccessporreponame:Repositório Institucional da UFMSinstname:Universidade Federal de Mato Grosso do Sul (UFMS)instacron:UFMSORIGINALtese_Nathália Fontoura_final.pdftese_Nathália Fontoura_final.pdfapplication/pdf1342555https://repositorio.ufms.br/bitstream/123456789/5389/-1/tese_Nath%c3%a1lia%20Fontoura_final.pdf5bcdb08f715894d265b04816265df844MD5-1123456789/53892022-11-30 15:18:42.262oai:repositorio.ufms.br:123456789/5389Repositório InstitucionalPUBhttps://repositorio.ufms.br/oai/requestri.prograd@ufms.bropendoar:21242022-11-30T19:18:42Repositório Institucional da UFMS - Universidade Federal de Mato Grosso do Sul (UFMS)false
dc.title.pt_BR.fl_str_mv AVALIAÇÃO DO DESEMPENHO DIAGNÓSTICO DE NOVAS TÉCNICAS SOROLÓGICAS PARA LEISHMANIOSE VISCERAL CANINA
title AVALIAÇÃO DO DESEMPENHO DIAGNÓSTICO DE NOVAS TÉCNICAS SOROLÓGICAS PARA LEISHMANIOSE VISCERAL CANINA
spellingShingle AVALIAÇÃO DO DESEMPENHO DIAGNÓSTICO DE NOVAS TÉCNICAS SOROLÓGICAS PARA LEISHMANIOSE VISCERAL CANINA
NATHALIA LOPES FONTOURA MATEUS
ELISA, fitas reagentes, proteína recombinante
proteína quimera, teste rápido
title_short AVALIAÇÃO DO DESEMPENHO DIAGNÓSTICO DE NOVAS TÉCNICAS SOROLÓGICAS PARA LEISHMANIOSE VISCERAL CANINA
title_full AVALIAÇÃO DO DESEMPENHO DIAGNÓSTICO DE NOVAS TÉCNICAS SOROLÓGICAS PARA LEISHMANIOSE VISCERAL CANINA
title_fullStr AVALIAÇÃO DO DESEMPENHO DIAGNÓSTICO DE NOVAS TÉCNICAS SOROLÓGICAS PARA LEISHMANIOSE VISCERAL CANINA
title_full_unstemmed AVALIAÇÃO DO DESEMPENHO DIAGNÓSTICO DE NOVAS TÉCNICAS SOROLÓGICAS PARA LEISHMANIOSE VISCERAL CANINA
title_sort AVALIAÇÃO DO DESEMPENHO DIAGNÓSTICO DE NOVAS TÉCNICAS SOROLÓGICAS PARA LEISHMANIOSE VISCERAL CANINA
author NATHALIA LOPES FONTOURA MATEUS
author_facet NATHALIA LOPES FONTOURA MATEUS
author_role author
dc.contributor.advisor1.fl_str_mv Carlos Alberto do Nascimento Ramos
dc.contributor.author.fl_str_mv NATHALIA LOPES FONTOURA MATEUS
contributor_str_mv Carlos Alberto do Nascimento Ramos
dc.subject.por.fl_str_mv ELISA, fitas reagentes, proteína recombinante
proteína quimera, teste rápido
topic ELISA, fitas reagentes, proteína recombinante
proteína quimera, teste rápido
description Visceral leishmaniasis (VL) is a zoonosis caused by the protozoan Leishmania infantum. Dogs are considered important urban reservoir for the parasite and canine visceral leishmaniasis (CVL) diagnosis contributes for monitoring and reemergence of VL. CVL confirmatory diagnosis as recommended by the Mistry of Health is performed using enzyme-linked immunosorbent assay (ELISA) with crude antigen of L. major-like. However, this technique may provide false positive results for animals infected with other trypanosomatids or Ehrlichia canis. Besides that, false negative results may be obtained for asymptomatic dogs, which can act as the parasite reservoir in the urban environment. In this context, recombinant antigens have been developed individually or combined to increase sensitivity, specificity, and accuracy of CVL serodiagnosis. This study analyzed three serologic tests for CVL diagnosis: two national developed ELISA using recombinant proteins (Q1 and Q5) and one rapid test using antigen of L. donovani promastigote membrane, in dipstick presentation. To evaluate chimeric proteins performances in ELISA, samples from a serological bank were used. Samples from naturally infected dogs with L. infantum symptomatic (n=30), oligosymptomatic (n=30) and asymptomatic (n=10); healthy (n=30), vaccinated with Leish-Tec (n=5) and Leishmune  (n=5) and infected with E. canis (n=30) were used. The performance of chimeric ELISAs were compared to the ELISA with L. infantum total extract as antigen. Greater values of sensitivity and specificity were observed, respectively, for Q1 (81.43%; 85.71%) and Q5 (74.29%; 90.0%), when compared to total extract antigen (71.43%; 82.86%). Although Q1 presented cross-reaction with Leish-Tec-vaccinated dogs, this chimeric protein demonstrated better sensitivity to diagnose asymptomatic (70%) and oligosymptomatic (73,33%) dogs. To access dipstick performance, samples from naturally infected dogs with L. infantum symptomatic (n=15), oligosymptomatic (n=15) and asymptomatic (n=9); healthy (n=15); vaccinated with Leish-Tec (n=5) and infected with E. canis (n=15) were obtained from a sera bank. The dipstick test presented higher sensitivity in the oligosymptomatic group (93.33%); followed by asymptomatic (77.88%) and symptomatic (73.33%); specificity value of 100% was observed for the test among all groups. Therefore, Q5 may be an interesting antigen for the development of serological tests and dipstick test could be a useful tool for CVL diagnosis due to its ease of performance. Both methods demonstrated good performance in situations where canine ehrlichiosis may be a differential diagnosis for the patient.
publishDate 2022
dc.date.accessioned.fl_str_mv 2022-11-30T19:18:41Z
dc.date.available.fl_str_mv 2022-11-30T19:18:41Z
dc.date.issued.fl_str_mv 2022
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
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dc.publisher.initials.fl_str_mv UFMS
dc.publisher.country.fl_str_mv Brasil
publisher.none.fl_str_mv Fundação Universidade Federal de Mato Grosso do Sul
dc.source.none.fl_str_mv reponame:Repositório Institucional da UFMS
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