Use of PCR–RFLP to identify Leishmania species in naturally-infected dogs.
Autor(a) principal: | |
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Data de Publicação: | 2006 |
Outros Autores: | , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UFOP |
Texto Completo: | http://www.repositorio.ufop.br/handle/123456789/1059 |
Resumo: | Tissue imprints on Giemsa stained slides from dogs were used to investigate the presence of Leishmania amastigotes by either optical microscopy (OM) or Polymerase chain reaction (PCR) detection of DNA. Samples from skin, spleen, lymph node, liver and bone marrow from a Leishmaniasis endemic area dogs where Leishmania ( Leishmania ) chagasi and Leishmania ( Viannia) braziliensis are sympatric were studied. Dogs were initially diagnosed by Indirect Immunofluorescence (IIF), as which 39 were IIF positive ( 1:40) and 16 negative. The IIF positive dogs were clinically grouped as symptomatic (n = 15), oligosymptomatic ( n = 12) and asymptomatic ( n = 12). Although PCR positivity was higher in symptomatic dogs, specially their skin samples, there was no significant difference among clinical groups or organs examined. Ten (62.5%) out of 16 IIF and OM negative animals were positive for PCR in at least one organ. Forty-eight positive PCR amplicons were further submitted to RFLP for Leishmania identification. All dogs were infected with L. ( L.) chagasiexcept one, infected withL. ( V.) braziliensis . PCR was more efficient than IIF and OM to diagnose canine visceral Leishmaniasis (CVL), regardless of the organ examined and the clinical form present. The use of PCR together with serology helps determining the extension of sub clinical infection in CVL endemic areas and provides a better estimate of the number of dogs to be targeted for control measures. In conclusion, our data reinforce the need for a specific diagnosis of canine infection in areas where diverse Leishmania species are sympatric and demonstrate that PCR–RFLP can be used to identify Leishmania species in dog tissue imprint stained slides. |
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Andrade, Hélida Monteiro deReis, Alexandre BarbosaSantos, Sara Lopes dosVolpini, Ângela CristinaMarques, Marcos JoséRomanha, Alvaro José2012-07-10T16:53:30Z2012-07-10T16:53:30Z2006ANDRADE, H. M. de et al. Use of PCR–RFLP to identify Leishmania species in naturally-infected dogs. Veterinary Parasitology, v. 140, n. 3-4, p. 231-238, set. 2006. Disponível em: <https://www.sciencedirect.com/science/article/pii/S0304401706002299>. Acesso em: 10 jul. 2012.03044017http://www.repositorio.ufop.br/handle/123456789/1059Tissue imprints on Giemsa stained slides from dogs were used to investigate the presence of Leishmania amastigotes by either optical microscopy (OM) or Polymerase chain reaction (PCR) detection of DNA. Samples from skin, spleen, lymph node, liver and bone marrow from a Leishmaniasis endemic area dogs where Leishmania ( Leishmania ) chagasi and Leishmania ( Viannia) braziliensis are sympatric were studied. Dogs were initially diagnosed by Indirect Immunofluorescence (IIF), as which 39 were IIF positive ( 1:40) and 16 negative. The IIF positive dogs were clinically grouped as symptomatic (n = 15), oligosymptomatic ( n = 12) and asymptomatic ( n = 12). Although PCR positivity was higher in symptomatic dogs, specially their skin samples, there was no significant difference among clinical groups or organs examined. Ten (62.5%) out of 16 IIF and OM negative animals were positive for PCR in at least one organ. Forty-eight positive PCR amplicons were further submitted to RFLP for Leishmania identification. All dogs were infected with L. ( L.) chagasiexcept one, infected withL. ( V.) braziliensis . PCR was more efficient than IIF and OM to diagnose canine visceral Leishmaniasis (CVL), regardless of the organ examined and the clinical form present. The use of PCR together with serology helps determining the extension of sub clinical infection in CVL endemic areas and provides a better estimate of the number of dogs to be targeted for control measures. In conclusion, our data reinforce the need for a specific diagnosis of canine infection in areas where diverse Leishmania species are sympatric and demonstrate that PCR–RFLP can be used to identify Leishmania species in dog tissue imprint stained slides.LeishmaniaIndentificationDogsDiagnosis of visceral leishmaniasisUse of PCR–RFLP to identify Leishmania species in naturally-infected dogs.info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleO periódico Veterinary Parasitology concede permissão para depósito deste artigo no Repositório Institucional da UFOP. Número da licença: 3291410216072.info:eu-repo/semantics/openAccessengreponame:Repositório Institucional da UFOPinstname:Universidade Federal de Ouro Preto (UFOP)instacron:UFOPLICENSElicense.txtlicense.txttext/plain; charset=utf-81748http://www.repositorio.ufop.br/bitstream/123456789/1059/5/license.txt8a4605be74aa9ea9d79846c1fba20a33MD55ORIGINALARTIGO_UseIdentifyLeishmania.pdfARTIGO_UseIdentifyLeishmania.pdfapplication/pdf145188http://www.repositorio.ufop.br/bitstream/123456789/1059/1/ARTIGO_UseIdentifyLeishmania.pdf21ca8b04354f7157f756e8feb5d08cdbMD51123456789/10592019-02-26 09:53:57.236oai:localhost: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Repositório InstitucionalPUBhttp://www.repositorio.ufop.br/oai/requestrepositorio@ufop.edu.bropendoar:32332019-02-26T14:53:57Repositório Institucional da UFOP - Universidade Federal de Ouro Preto (UFOP)false |
dc.title.pt_BR.fl_str_mv |
Use of PCR–RFLP to identify Leishmania species in naturally-infected dogs. |
title |
Use of PCR–RFLP to identify Leishmania species in naturally-infected dogs. |
spellingShingle |
Use of PCR–RFLP to identify Leishmania species in naturally-infected dogs. Andrade, Hélida Monteiro de Leishmania Indentification Dogs Diagnosis of visceral leishmaniasis |
title_short |
Use of PCR–RFLP to identify Leishmania species in naturally-infected dogs. |
title_full |
Use of PCR–RFLP to identify Leishmania species in naturally-infected dogs. |
title_fullStr |
Use of PCR–RFLP to identify Leishmania species in naturally-infected dogs. |
title_full_unstemmed |
Use of PCR–RFLP to identify Leishmania species in naturally-infected dogs. |
title_sort |
Use of PCR–RFLP to identify Leishmania species in naturally-infected dogs. |
author |
Andrade, Hélida Monteiro de |
author_facet |
Andrade, Hélida Monteiro de Reis, Alexandre Barbosa Santos, Sara Lopes dos Volpini, Ângela Cristina Marques, Marcos José Romanha, Alvaro José |
author_role |
author |
author2 |
Reis, Alexandre Barbosa Santos, Sara Lopes dos Volpini, Ângela Cristina Marques, Marcos José Romanha, Alvaro José |
author2_role |
author author author author author |
dc.contributor.author.fl_str_mv |
Andrade, Hélida Monteiro de Reis, Alexandre Barbosa Santos, Sara Lopes dos Volpini, Ângela Cristina Marques, Marcos José Romanha, Alvaro José |
dc.subject.por.fl_str_mv |
Leishmania Indentification Dogs Diagnosis of visceral leishmaniasis |
topic |
Leishmania Indentification Dogs Diagnosis of visceral leishmaniasis |
description |
Tissue imprints on Giemsa stained slides from dogs were used to investigate the presence of Leishmania amastigotes by either optical microscopy (OM) or Polymerase chain reaction (PCR) detection of DNA. Samples from skin, spleen, lymph node, liver and bone marrow from a Leishmaniasis endemic area dogs where Leishmania ( Leishmania ) chagasi and Leishmania ( Viannia) braziliensis are sympatric were studied. Dogs were initially diagnosed by Indirect Immunofluorescence (IIF), as which 39 were IIF positive ( 1:40) and 16 negative. The IIF positive dogs were clinically grouped as symptomatic (n = 15), oligosymptomatic ( n = 12) and asymptomatic ( n = 12). Although PCR positivity was higher in symptomatic dogs, specially their skin samples, there was no significant difference among clinical groups or organs examined. Ten (62.5%) out of 16 IIF and OM negative animals were positive for PCR in at least one organ. Forty-eight positive PCR amplicons were further submitted to RFLP for Leishmania identification. All dogs were infected with L. ( L.) chagasiexcept one, infected withL. ( V.) braziliensis . PCR was more efficient than IIF and OM to diagnose canine visceral Leishmaniasis (CVL), regardless of the organ examined and the clinical form present. The use of PCR together with serology helps determining the extension of sub clinical infection in CVL endemic areas and provides a better estimate of the number of dogs to be targeted for control measures. In conclusion, our data reinforce the need for a specific diagnosis of canine infection in areas where diverse Leishmania species are sympatric and demonstrate that PCR–RFLP can be used to identify Leishmania species in dog tissue imprint stained slides. |
publishDate |
2006 |
dc.date.issued.fl_str_mv |
2006 |
dc.date.accessioned.fl_str_mv |
2012-07-10T16:53:30Z |
dc.date.available.fl_str_mv |
2012-07-10T16:53:30Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
ANDRADE, H. M. de et al. Use of PCR–RFLP to identify Leishmania species in naturally-infected dogs. Veterinary Parasitology, v. 140, n. 3-4, p. 231-238, set. 2006. Disponível em: <https://www.sciencedirect.com/science/article/pii/S0304401706002299>. Acesso em: 10 jul. 2012. |
dc.identifier.uri.fl_str_mv |
http://www.repositorio.ufop.br/handle/123456789/1059 |
dc.identifier.issn.none.fl_str_mv |
03044017 |
identifier_str_mv |
ANDRADE, H. M. de et al. Use of PCR–RFLP to identify Leishmania species in naturally-infected dogs. Veterinary Parasitology, v. 140, n. 3-4, p. 231-238, set. 2006. Disponível em: <https://www.sciencedirect.com/science/article/pii/S0304401706002299>. Acesso em: 10 jul. 2012. 03044017 |
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http://www.repositorio.ufop.br/handle/123456789/1059 |
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