Anaplasma platys e Ehrlichia canis em cães: avaliação de alterações oculares, desenvolvimento e validação de técnica de diagnóstico molecular

Detalhes bibliográficos
Autor(a) principal: Costa, Hérika Xavier da
Data de Publicação: 2015
Tipo de documento: Tese
Idioma: por
Título da fonte: Repositório Institucional da UFG
Texto Completo: http://repositorio.bc.ufg.br/tede/handle/tede/5850
Resumo: Anaplasma platys and Ehrlichia canis are obligate intracellular rickettsial organisms of dogs. A. platys appear to parasitize only platelets causing canine infectious cyclic thrombocytopenia (CICT), whereas E. canis has tropism for circulating mononuclear cells causing canine monocytic ehrlichiosis (CME). Clinical signs of CME and CICT are nonspecific and common to many other diseases transmitted by ticks. Definitive diagnosis of A. platys and E. canis is made by specific diagnostic tests; however, two new real time PCR protocols (qPCR) for the detection of A. platys and E. canis were developed from reference sequences for the 16S rRNA gene of A. platys and E. canis. New primers and probes were able to detect A. platys and E. canis. Identification of natural infections in dogs by A. platys and E. canis, previously confirmed by conventional PCR, showed high correlation sensitivity (100% for A. platys and E. canis) and epidemiological specificity (100% for A. platys and 98,41% for E. canis) with new qPCR using hydrolyses probe (TaqMan) protocols Analytical specificity tests produced species-specific results, not occurring DNA amplification of the following blood parasites Babesia vogeli, Hepatozoon canis and Mycoplasma haemofelis. R. sanguineus DNA. Two new qPCR protocols described were able to detect 0,14 fg of E. canis DNA and 0,9 fg of A. platys DNA and represent alternative as a diagnostic tool for the specific detection of A. platys and E. canis being useful for diagnosis of these canine hemoparasitosis. Ocular abnormalities are among the clinical signs found in dogs infected with E. canis and cases of uveitis have been attributed to infection with A. platys, but the frequency of eye injuries in cases of infection with A. platys is still unknown. After searching the frequency of infections by A. platys and E. canis in 100 dogs with ocular (OA) and 100 dogs without ocular alterations (WOA), using the PCR as a diagnostic method, we found that among the hemoparasites, A. platys and E. canis detected and identified by PCR, E. canis was the most frequent, both in the group of OA dogs (33%; 33/100) and in the group of WOA dogs (24%; 24 / 100). The frequency of A. platys was 5% (5/100) for the group of OA dogs and 4% (4/100) for the group of WOA dogs. Ocular changes associated with infection by E. canis were bilateral uveitis and retinal detachment. It was not possible to verify the relationship of ocular manifestations with infection by A. platys due to the low number of dogs with ocular abnormalities infected with this rickettsia.
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spelling Linhares, Guido Fontgalland Coelhohttp://lattes.cnpq.br/6261928164195145Duarte, Sabrina Castilhohttp://lattes.cnpq.br/4703399111676562Jayme, Valéria de Sáhttp://lattes.cnpq.br/0603234425928309Linhares, Guido Fontgalland CoelhoMassard, Carlos LuizFernandes, Osvaldo Kort KampOliveira, Vanessa Silvestre Ferreira dehttp://lattes.cnpq.br/9319333962450391Costa, Hérika Xavier da2016-08-05T15:01:18Z2015-04-23COSTA, H. X. Anaplasma platys e Ehrlichia canis em cães: avaliação de alterações oculares, desenvolvimento e validação de técnica de diagnóstico molecular. 2015. 60 f. Tese (Doutorado em Ciência Animal) - Universidade Federal de Goiás, Goiânia, 2015.http://repositorio.bc.ufg.br/tede/handle/tede/5850Anaplasma platys and Ehrlichia canis are obligate intracellular rickettsial organisms of dogs. A. platys appear to parasitize only platelets causing canine infectious cyclic thrombocytopenia (CICT), whereas E. canis has tropism for circulating mononuclear cells causing canine monocytic ehrlichiosis (CME). Clinical signs of CME and CICT are nonspecific and common to many other diseases transmitted by ticks. Definitive diagnosis of A. platys and E. canis is made by specific diagnostic tests; however, two new real time PCR protocols (qPCR) for the detection of A. platys and E. canis were developed from reference sequences for the 16S rRNA gene of A. platys and E. canis. New primers and probes were able to detect A. platys and E. canis. Identification of natural infections in dogs by A. platys and E. canis, previously confirmed by conventional PCR, showed high correlation sensitivity (100% for A. platys and E. canis) and epidemiological specificity (100% for A. platys and 98,41% for E. canis) with new qPCR using hydrolyses probe (TaqMan) protocols Analytical specificity tests produced species-specific results, not occurring DNA amplification of the following blood parasites Babesia vogeli, Hepatozoon canis and Mycoplasma haemofelis. R. sanguineus DNA. Two new qPCR protocols described were able to detect 0,14 fg of E. canis DNA and 0,9 fg of A. platys DNA and represent alternative as a diagnostic tool for the specific detection of A. platys and E. canis being useful for diagnosis of these canine hemoparasitosis. Ocular abnormalities are among the clinical signs found in dogs infected with E. canis and cases of uveitis have been attributed to infection with A. platys, but the frequency of eye injuries in cases of infection with A. platys is still unknown. After searching the frequency of infections by A. platys and E. canis in 100 dogs with ocular (OA) and 100 dogs without ocular alterations (WOA), using the PCR as a diagnostic method, we found that among the hemoparasites, A. platys and E. canis detected and identified by PCR, E. canis was the most frequent, both in the group of OA dogs (33%; 33/100) and in the group of WOA dogs (24%; 24 / 100). The frequency of A. platys was 5% (5/100) for the group of OA dogs and 4% (4/100) for the group of WOA dogs. Ocular changes associated with infection by E. canis were bilateral uveitis and retinal detachment. It was not possible to verify the relationship of ocular manifestations with infection by A. platys due to the low number of dogs with ocular abnormalities infected with this rickettsia.Anaplasma platys e Ehrlichia canis são riquétsias intracelulares obrigatórias de cães. A. platys parasita plaquetas causando a trombocitopenia cíclica infecciosa canina (TCIC), enquanto que E. canis possui tropismo por células mononucleares circulantes, causando a erliquiose monocítica canina (EMC). Os sinais clínicos da EMC e TCIC são inespecíficos e comuns a inúmeras outras enfermidades transmitidas por carrapatos. O diagnóstico definitivo de E. canis e A. platys é firmado através do emprego de testes de diagnóstico específicos. Dessa forma, dois novos protocolos de ensaios de PCR em tempo real (qPCR) para o detecção de A. platys e E. canis foram desenvolvidos a partir de sequências de referência para o gene 16S rRNA de A. platys e E. canis. Os novos iniciadores e sondas desenhados foram capazes de detectar A. platys e E. canis. Identificação de infecções naturais em cães por A. platys e E. canis, previamente confirmada por PCR convencional demonstrou alta correlação na sensibilidade (100% para A. platys e E. canis) e especificidade epidemiológicas (100% para A. platys e 98,41% para E. canis) com os novos protocolos de qPCR utilizando sondas de hidrólise TaqMan. Os testes de especificidade analítica produziram resultado espécie específicos, não ocorrendo amplificação de DNA dos seguintes hemoparasitos: Babesia vogeli, Hepatozoon canis e Mycoplasma haemofelis. DNA de Rhipicephalus sanguineus também não foi amplificado. Os dois novos protocolos de qPCR descritos aqui foram capazes de detectar até 0,14 fg de DNA de E. canis e 0,9 fg de DNA de A. platys e representam alternativas como ferramenta de diagnóstico para a detecção específica de A. platys e E. canis, sendo útil para o diagnóstico dessas hemoparasitoses em cães. As alterações oculares estão entre os sinais clínicos encontrados em cães infectados com Ehrlichia canis e casos de uveítes já foram atribuídos a infecção por A. platys, porém a frequência de lesões oculares em casos de infecção por A. platys ainda é desconhecida. Após pesquisar a frequência de infecções por E. canis e A. platys em 100 cães com alterações oculares (CAO) e 100 cães sem alterações oculares (SAO), utilizando-se a PCR como método de diagnóstico, verificou-se que dentre os hemoparasitos, E. canis e A. platys, detectados e identificados por PCR, E. canis foi o de maior frequência, tanto no grupo de cães CAO (33%; 33/100) quanto no grupo de cães SAO (24%; 24/100). A frequência de A. platys foi de 5% (5/100) para o grupo de cães CAO e 4% (4/100) para o grupo de cães SAO. As alterações oculares que apresentaram relação com a infecção por E. canis foram uveíte bilateral e descolamento de retina. Não foi possível verificar a relação das alterações oculares com a infecção por A. platys devido ao baixo número de cães com alterações oculares infectados com esta riquétsia.Submitted by Cássia Santos (cassia.bcufg@gmail.com) on 2016-08-05T10:36:05Z No. of bitstreams: 2 Tese - Herika Xavier da Costa - 2015.pdf: 2103590 bytes, checksum: f8dd254901af7084719caef9647c0af2 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-08-05T15:01:18Z (GMT) No. of bitstreams: 2 Tese - Herika Xavier da Costa - 2015.pdf: 2103590 bytes, checksum: f8dd254901af7084719caef9647c0af2 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)Made available in DSpace on 2016-08-05T15:01:18Z (GMT). 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dc.title.por.fl_str_mv Anaplasma platys e Ehrlichia canis em cães: avaliação de alterações oculares, desenvolvimento e validação de técnica de diagnóstico molecular
dc.title.alternative.eng.fl_str_mv Anaplasma platys and Ehrlichia canis in dogs: ocular evaluation development and validation of molecular diagnostic technical
title Anaplasma platys e Ehrlichia canis em cães: avaliação de alterações oculares, desenvolvimento e validação de técnica de diagnóstico molecular
spellingShingle Anaplasma platys e Ehrlichia canis em cães: avaliação de alterações oculares, desenvolvimento e validação de técnica de diagnóstico molecular
Costa, Hérika Xavier da
Hemoparasitoses canina
Cães
Diagnóstico molecular
PCR
Canine hemoparasitosis
Dogs
Molecular diagnosis
PCR
MEDICINA VETERINARIA PREVENTIVA::SAUDE ANIMAL (PROGRAMAS SANITARIOS)
title_short Anaplasma platys e Ehrlichia canis em cães: avaliação de alterações oculares, desenvolvimento e validação de técnica de diagnóstico molecular
title_full Anaplasma platys e Ehrlichia canis em cães: avaliação de alterações oculares, desenvolvimento e validação de técnica de diagnóstico molecular
title_fullStr Anaplasma platys e Ehrlichia canis em cães: avaliação de alterações oculares, desenvolvimento e validação de técnica de diagnóstico molecular
title_full_unstemmed Anaplasma platys e Ehrlichia canis em cães: avaliação de alterações oculares, desenvolvimento e validação de técnica de diagnóstico molecular
title_sort Anaplasma platys e Ehrlichia canis em cães: avaliação de alterações oculares, desenvolvimento e validação de técnica de diagnóstico molecular
author Costa, Hérika Xavier da
author_facet Costa, Hérika Xavier da
author_role author
dc.contributor.advisor1.fl_str_mv Linhares, Guido Fontgalland Coelho
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/6261928164195145
dc.contributor.advisor-co1.fl_str_mv Duarte, Sabrina Castilho
dc.contributor.advisor-co1Lattes.fl_str_mv http://lattes.cnpq.br/4703399111676562
dc.contributor.advisor-co2.fl_str_mv Jayme, Valéria de Sá
dc.contributor.advisor-co2Lattes.fl_str_mv http://lattes.cnpq.br/0603234425928309
dc.contributor.referee1.fl_str_mv Linhares, Guido Fontgalland Coelho
dc.contributor.referee2.fl_str_mv Massard, Carlos Luiz
dc.contributor.referee3.fl_str_mv Fernandes, Osvaldo Kort Kamp
dc.contributor.referee4.fl_str_mv Oliveira, Vanessa Silvestre Ferreira de
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/9319333962450391
dc.contributor.author.fl_str_mv Costa, Hérika Xavier da
contributor_str_mv Linhares, Guido Fontgalland Coelho
Duarte, Sabrina Castilho
Jayme, Valéria de Sá
Linhares, Guido Fontgalland Coelho
Massard, Carlos Luiz
Fernandes, Osvaldo Kort Kamp
Oliveira, Vanessa Silvestre Ferreira de
dc.subject.por.fl_str_mv Hemoparasitoses canina
Cães
Diagnóstico molecular
PCR
topic Hemoparasitoses canina
Cães
Diagnóstico molecular
PCR
Canine hemoparasitosis
Dogs
Molecular diagnosis
PCR
MEDICINA VETERINARIA PREVENTIVA::SAUDE ANIMAL (PROGRAMAS SANITARIOS)
dc.subject.eng.fl_str_mv Canine hemoparasitosis
Dogs
Molecular diagnosis
PCR
dc.subject.cnpq.fl_str_mv MEDICINA VETERINARIA PREVENTIVA::SAUDE ANIMAL (PROGRAMAS SANITARIOS)
description Anaplasma platys and Ehrlichia canis are obligate intracellular rickettsial organisms of dogs. A. platys appear to parasitize only platelets causing canine infectious cyclic thrombocytopenia (CICT), whereas E. canis has tropism for circulating mononuclear cells causing canine monocytic ehrlichiosis (CME). Clinical signs of CME and CICT are nonspecific and common to many other diseases transmitted by ticks. Definitive diagnosis of A. platys and E. canis is made by specific diagnostic tests; however, two new real time PCR protocols (qPCR) for the detection of A. platys and E. canis were developed from reference sequences for the 16S rRNA gene of A. platys and E. canis. New primers and probes were able to detect A. platys and E. canis. Identification of natural infections in dogs by A. platys and E. canis, previously confirmed by conventional PCR, showed high correlation sensitivity (100% for A. platys and E. canis) and epidemiological specificity (100% for A. platys and 98,41% for E. canis) with new qPCR using hydrolyses probe (TaqMan) protocols Analytical specificity tests produced species-specific results, not occurring DNA amplification of the following blood parasites Babesia vogeli, Hepatozoon canis and Mycoplasma haemofelis. R. sanguineus DNA. Two new qPCR protocols described were able to detect 0,14 fg of E. canis DNA and 0,9 fg of A. platys DNA and represent alternative as a diagnostic tool for the specific detection of A. platys and E. canis being useful for diagnosis of these canine hemoparasitosis. Ocular abnormalities are among the clinical signs found in dogs infected with E. canis and cases of uveitis have been attributed to infection with A. platys, but the frequency of eye injuries in cases of infection with A. platys is still unknown. After searching the frequency of infections by A. platys and E. canis in 100 dogs with ocular (OA) and 100 dogs without ocular alterations (WOA), using the PCR as a diagnostic method, we found that among the hemoparasites, A. platys and E. canis detected and identified by PCR, E. canis was the most frequent, both in the group of OA dogs (33%; 33/100) and in the group of WOA dogs (24%; 24 / 100). The frequency of A. platys was 5% (5/100) for the group of OA dogs and 4% (4/100) for the group of WOA dogs. Ocular changes associated with infection by E. canis were bilateral uveitis and retinal detachment. It was not possible to verify the relationship of ocular manifestations with infection by A. platys due to the low number of dogs with ocular abnormalities infected with this rickettsia.
publishDate 2015
dc.date.issued.fl_str_mv 2015-04-23
dc.date.accessioned.fl_str_mv 2016-08-05T15:01:18Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/doctoralThesis
format doctoralThesis
status_str publishedVersion
dc.identifier.citation.fl_str_mv COSTA, H. X. Anaplasma platys e Ehrlichia canis em cães: avaliação de alterações oculares, desenvolvimento e validação de técnica de diagnóstico molecular. 2015. 60 f. Tese (Doutorado em Ciência Animal) - Universidade Federal de Goiás, Goiânia, 2015.
dc.identifier.uri.fl_str_mv http://repositorio.bc.ufg.br/tede/handle/tede/5850
identifier_str_mv COSTA, H. X. Anaplasma platys e Ehrlichia canis em cães: avaliação de alterações oculares, desenvolvimento e validação de técnica de diagnóstico molecular. 2015. 60 f. Tese (Doutorado em Ciência Animal) - Universidade Federal de Goiás, Goiânia, 2015.
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