Desempenho da técnica de citometria de fluxo, no diagnóstico precoce da toxoplasmose congênita

Detalhes bibliográficos
Autor(a) principal: Tonini, Aline de Castro Zacche
Data de Publicação: 2014
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da Universidade Federal do Espírito Santo (riUfes)
Texto Completo: http://repositorio.ufes.br/handle/10/1604
Resumo: The routine diagnosis of pos-natal congenital toxoplasmosis is based on the detection of IgM and/or IgA anti-T. gondii antibodies in neonates sera. However, in a significant number of infected newborns these antibodies are not detected, hindering the early diagnosis of the disease. Therefore, in this study it was evaluated the performance of the detection of anti-T. gondii antibodies by flow cytometry in the sorological diagnosis of early pos-natal congenital toxoplasmosis., Eight eight sera samples of children with congenital infection by T.gondii (TOXO) and 19 sera samples of uninfected children were evaluated. The sensitivity of the test was 47.6% for IgM, 72.6% for IgA, and 75% for IgG, with 100% of specificity for all tests. As to the IgG subclasses, the sensitivity was 73.9% for IgG1, 60.2% for IgG2, and 83% for IgG3 with 100% of specificity for all tests. The sensitivity of IgG4 was superior to the aformentioned subclasses reaching 94.7%, although achieved a poor specificity of 4.6%. The IgG avidity when employed to segregate TOXO and NI groups presented a great performance, with 97% of sensitivity and 93% of specificity. It was also done a comparative analysis of the presence of IgG and IgG3 in children of TOXO and NI groups with their respective mothers. Our results showed that the children with congenital toxoplasmosis presented an average reactivity of these antibodies equivalent to their respective mothers, which suggests that they are producing antibodies in response to T.gondii infection. In contrast, uninfected children presented a lower average reactivity of those antibodies in comparison to their respective mothers. In addition, our study proposed an algorithm for the diagnosis of congenital toxoplasmosis using IgM test performed by conventional methods available in routine clinical laboratories as the initial approach followed by the search for IgG3 and IgG avidity using flow cytometry, reaching a final performance of 98% of sensitivity and 93% of specificity. Data analysis showed the applicability of flow cytometry for the detection of IgG3 antibodies and IgG anti-T. gondii avidity as a complementary tool for the early diagnosis of congenital toxoplasmosis.
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spelling Pereira, Fausto Edmundo LimaLemos, Elenice MoreiraTonini, Aline de Castro ZaccheBarros, Geisa BaptistaFux, Blima2015-11-16T21:07:49Z2016-06-24T06:00:07Z2014-12-042014-12-04The routine diagnosis of pos-natal congenital toxoplasmosis is based on the detection of IgM and/or IgA anti-T. gondii antibodies in neonates sera. However, in a significant number of infected newborns these antibodies are not detected, hindering the early diagnosis of the disease. Therefore, in this study it was evaluated the performance of the detection of anti-T. gondii antibodies by flow cytometry in the sorological diagnosis of early pos-natal congenital toxoplasmosis., Eight eight sera samples of children with congenital infection by T.gondii (TOXO) and 19 sera samples of uninfected children were evaluated. The sensitivity of the test was 47.6% for IgM, 72.6% for IgA, and 75% for IgG, with 100% of specificity for all tests. As to the IgG subclasses, the sensitivity was 73.9% for IgG1, 60.2% for IgG2, and 83% for IgG3 with 100% of specificity for all tests. The sensitivity of IgG4 was superior to the aformentioned subclasses reaching 94.7%, although achieved a poor specificity of 4.6%. The IgG avidity when employed to segregate TOXO and NI groups presented a great performance, with 97% of sensitivity and 93% of specificity. It was also done a comparative analysis of the presence of IgG and IgG3 in children of TOXO and NI groups with their respective mothers. Our results showed that the children with congenital toxoplasmosis presented an average reactivity of these antibodies equivalent to their respective mothers, which suggests that they are producing antibodies in response to T.gondii infection. In contrast, uninfected children presented a lower average reactivity of those antibodies in comparison to their respective mothers. In addition, our study proposed an algorithm for the diagnosis of congenital toxoplasmosis using IgM test performed by conventional methods available in routine clinical laboratories as the initial approach followed by the search for IgG3 and IgG avidity using flow cytometry, reaching a final performance of 98% of sensitivity and 93% of specificity. Data analysis showed the applicability of flow cytometry for the detection of IgG3 antibodies and IgG anti-T. gondii avidity as a complementary tool for the early diagnosis of congenital toxoplasmosis.O diagnóstico de rotina pós-natal da toxoplasmose congênita é baseado na detecção de anticorpos IgM e/ou IgA anti-T. gondii no soro dos neonatos. No entanto, em um número significativo de neonatos infectados esses anticorpos não são detectados, dificultando o diagnóstico precoce da doença. Portanto, neste estudo foi avaliado o desempenho da pesquisa de anticorpos anti-T. gondii por citometria de fluxo no diagnóstico sorológico pós-natal precoce da toxoplasmose congênita. Foram avaliados 88 amostras de soro de crianças com infecção congênita pelo T. gondii (TOXO) e 19 amostras de soro de crianças não infectadas (NI). A sensibilidade do teste foi de 47,6% para IgM, 72,6% para IgA e 75% para IgG, com 100% de especificidade para todos os testes. Quanto a pesquisa de subclasses de IgG, a sensibilidade foi de 73,9% para IgG1, 60,2% para IgG2 e 83% para IgG3 com 100% de especificidade para todos os testes. A sensibilidade de IgG4 foi superior às demais, alcançando 94,7%, embora apresentou uma baixa especificidade de 4,6%. A pesquisa da avidez de IgG quando aplicada para segregação dos grupos TOXO e NI, apresentou um ótimo desempenho com 97% de sensibilidade e 93% de especificidade. Foi também realizado uma análise comparativa da presença de IgG e IgG3 em crianças do grupo TOXO e NI com suas respectivas mães. Nossos resultados mostraram que as crianças infectadas apresentaram uma reatividade média desses anticorpos equivalentes a de suas respectivas mães, o que sugere estarem sintetizando estes anticorpos em resposta a infecção pelo T. gondii. Em contraste, as crianças não infectadas apresentaram uma menor reatividade média desses anticorpos em relação à suas respectivas mães. Além disso, nosso estudo propôs um algoritmo para o diagnóstico da toxoplasmose congênita utilizando o teste de IgM por métodos convencionais disponíveis na rotina dos laboratórios clínicos como estratégia inicial seguido da pesquisa de IgG3 e avidez de IgG pela citometria de fluxo com desempenho final de 98% de sensibilidade e 93% de especificidade. A análise dos dados demonstrou a aplicabilidade da citometria de fluxo para a pesquisa de anticorpos IgG3 e avidez de IgG anti-T. gondii como uma ferramenta complementar para o diagnóstico precoce da toxoplasmose congênita.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESTexthttp://repositorio.ufes.br/handle/10/1604porUniversidade Federal do Espírito SantoMestrado em Doenças InfecciosasPrograma de Pós-Graduação em Doenças InfecciosasUFESBRCentro de Ciências da SaúdeToxoplasmose congênitaCitometria de fluxoDiagnósticoDiagnóstico sorológicoDoenças Infecciosas e Parasitárias61Desempenho da técnica de citometria de fluxo, no diagnóstico precoce da toxoplasmose congênitainfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da Universidade Federal do Espírito Santo (riUfes)instname:Universidade Federal do Espírito Santo (UFES)instacron:UFESCoordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESORIGINALDESEMPENHO DA TÉCNICA DE CITOMETRIA DE FLUXO, NO.pdfDESEMPENHO DA TÉCNICA DE CITOMETRIA DE FLUXO, NO.pdfapplication/pdf3747193http://repositorio.ufes.br/bitstreams/6be668a6-fd21-48f8-8f8a-ad94d1499e64/downloadef571a9dce24b87b45d8fd8d89916048MD51CC-LICENSElicense_urllicense_urltext/plain; charset=utf-849http://repositorio.ufes.br/bitstreams/31da8240-5acc-43af-934a-3c3ddcaf382b/download4afdbb8c545fd630ea7db775da747b2fMD52license_textlicense_texttext/html; charset=utf-821468http://repositorio.ufes.br/bitstreams/ee0b5be1-1292-4431-bad0-ad674d2afae7/downloadae2fe251842ade1134c5d9bb99b6eefeMD53license_rdflicense_rdfapplication/rdf+xml; charset=utf-823148http://repositorio.ufes.br/bitstreams/1d39bd96-9d6c-44d8-adb9-19722dea8838/download9da0b6dfac957114c6a7714714b86306MD54LICENSElicense.txtlicense.txttext/plain; charset=utf-81748http://repositorio.ufes.br/bitstreams/1b52ebe7-6531-4c8e-8817-99b4746c9e54/download8a4605be74aa9ea9d79846c1fba20a33MD5510/16042024-07-16 17:04:09.224oai:repositorio.ufes.br:10/1604http://repositorio.ufes.brRepositório InstitucionalPUBhttp://repositorio.ufes.br/oai/requestopendoar:21082024-10-15T17:53:04.949290Repositório Institucional da Universidade Federal do Espírito Santo (riUfes) - Universidade Federal do Espírito Santo (UFES)falseTk9URTogUExBQ0UgWU9VUiBPV04gTElDRU5TRSBIRVJFClRoaXMgc2FtcGxlIGxpY2Vuc2UgaXMgcHJvdmlkZWQgZm9yIGluZm9ybWF0aW9uYWwgcHVycG9zZXMgb25seS4KCk5PTi1FWENMVVNJVkUgRElTVFJJQlVUSU9OIExJQ0VOU0UKCkJ5IHNpZ25pbmcgYW5kIHN1Ym1pdHRpbmcgdGhpcyBsaWNlbnNlLCB5b3UgKHRoZSBhdXRob3Iocykgb3IgY29weXJpZ2h0Cm93bmVyKSBncmFudHMgdG8gRFNwYWNlIFVuaXZlcnNpdHkgKERTVSkgdGhlIG5vbi1leGNsdXNpdmUgcmlnaHQgdG8gcmVwcm9kdWNlLAp0cmFuc2xhdGUgKGFzIGRlZmluZWQgYmVsb3cpLCBhbmQvb3IgZGlzdHJpYnV0ZSB5b3VyIHN1Ym1pc3Npb24gKGluY2x1ZGluZwp0aGUgYWJzdHJhY3QpIHdvcmxkd2lkZSBpbiBwcmludCBhbmQgZWxlY3Ryb25pYyBmb3JtYXQgYW5kIGluIGFueSBtZWRpdW0sCmluY2x1ZGluZyBidXQgbm90IGxpbWl0ZWQgdG8gYXVkaW8gb3IgdmlkZW8uCgpZb3UgYWdyZWUgdGhhdCBEU1UgbWF5LCB3aXRob3V0IGNoYW5naW5nIHRoZSBjb250ZW50LCB0cmFuc2xhdGUgdGhlCnN1Ym1pc3Npb24gdG8gYW55IG1lZGl1bSBvciBmb3JtYXQgZm9yIHRoZSBwdXJwb3NlIG9mIHByZXNlcnZhdGlvbi4KCllvdSBhbHNvIGFncmVlIHRoYXQgRFNVIG1heSBrZWVwIG1vcmUgdGhhbiBvbmUgY29weSBvZiB0aGlzIHN1Ym1pc3Npb24gZm9yCnB1cnBvc2VzIG9mIHNlY3VyaXR5LCBiYWNrLXVwIGFuZCBwcmVzZXJ2YXRpb24uCgpZb3UgcmVwcmVzZW50IHRoYXQgdGhlIHN1Ym1pc3Npb24gaXMgeW91ciBvcmlnaW5hbCB3b3JrLCBhbmQgdGhhdCB5b3UgaGF2ZQp0aGUgcmlnaHQgdG8gZ3JhbnQgdGhlIHJpZ2h0cyBjb250YWluZWQgaW4gdGhpcyBsaWNlbnNlLiBZb3UgYWxzbyByZXByZXNlbnQKdGhhdCB5b3VyIHN1Ym1pc3Npb24gZG9lcyBub3QsIHRvIHRoZSBiZXN0IG9mIHlvdXIga25vd2xlZGdlLCBpbmZyaW5nZSB1cG9uCmFueW9uZSdzIGNvcHlyaWdodC4KCklmIHRoZSBzdWJtaXNzaW9uIGNvbnRhaW5zIG1hdGVyaWFsIGZvciB3aGljaCB5b3UgZG8gbm90IGhvbGQgY29weXJpZ2h0LAp5b3UgcmVwcmVzZW50IHRoYXQgeW91IGhhdmUgb2J0YWluZWQgdGhlIHVucmVzdHJpY3RlZCBwZXJtaXNzaW9uIG9mIHRoZQpjb3B5cmlnaHQgb3duZXIgdG8gZ3JhbnQgRFNVIHRoZSByaWdodHMgcmVxdWlyZWQgYnkgdGhpcyBsaWNlbnNlLCBhbmQgdGhhdApzdWNoIHRoaXJkLXBhcnR5IG93bmVkIG1hdGVyaWFsIGlzIGNsZWFybHkgaWRlbnRpZmllZCBhbmQgYWNrbm93bGVkZ2VkCndpdGhpbiB0aGUgdGV4dCBvciBjb250ZW50IG9mIHRoZSBzdWJtaXNzaW9uLgoKSUYgVEhFIFNVQk1JU1NJT04gSVMgQkFTRUQgVVBPTiBXT1JLIFRIQVQgSEFTIEJFRU4gU1BPTlNPUkVEIE9SIFNVUFBPUlRFRApCWSBBTiBBR0VOQ1kgT1IgT1JHQU5JWkFUSU9OIE9USEVSIFRIQU4gRFNVLCBZT1UgUkVQUkVTRU5UIFRIQVQgWU9VIEhBVkUKRlVMRklMTEVEIEFOWSBSSUdIVCBPRiBSRVZJRVcgT1IgT1RIRVIgT0JMSUdBVElPTlMgUkVRVUlSRUQgQlkgU1VDSApDT05UUkFDVCBPUiBBR1JFRU1FTlQuCgpEU1Ugd2lsbCBjbGVhcmx5IGlkZW50aWZ5IHlvdXIgbmFtZShzKSBhcyB0aGUgYXV0aG9yKHMpIG9yIG93bmVyKHMpIG9mIHRoZQpzdWJtaXNzaW9uLCBhbmQgd2lsbCBub3QgbWFrZSBhbnkgYWx0ZXJhdGlvbiwgb3RoZXIgdGhhbiBhcyBhbGxvd2VkIGJ5IHRoaXMKbGljZW5zZSwgdG8geW91ciBzdWJtaXNzaW9uLgo=
dc.title.none.fl_str_mv Desempenho da técnica de citometria de fluxo, no diagnóstico precoce da toxoplasmose congênita
title Desempenho da técnica de citometria de fluxo, no diagnóstico precoce da toxoplasmose congênita
spellingShingle Desempenho da técnica de citometria de fluxo, no diagnóstico precoce da toxoplasmose congênita
Tonini, Aline de Castro Zacche
Toxoplasmose congênita
Citometria de fluxo
Diagnóstico
Diagnóstico sorológico
Doenças Infecciosas e Parasitárias
61
title_short Desempenho da técnica de citometria de fluxo, no diagnóstico precoce da toxoplasmose congênita
title_full Desempenho da técnica de citometria de fluxo, no diagnóstico precoce da toxoplasmose congênita
title_fullStr Desempenho da técnica de citometria de fluxo, no diagnóstico precoce da toxoplasmose congênita
title_full_unstemmed Desempenho da técnica de citometria de fluxo, no diagnóstico precoce da toxoplasmose congênita
title_sort Desempenho da técnica de citometria de fluxo, no diagnóstico precoce da toxoplasmose congênita
author Tonini, Aline de Castro Zacche
author_facet Tonini, Aline de Castro Zacche
author_role author
dc.contributor.advisor-co1.fl_str_mv Pereira, Fausto Edmundo Lima
dc.contributor.advisor1.fl_str_mv Lemos, Elenice Moreira
dc.contributor.author.fl_str_mv Tonini, Aline de Castro Zacche
dc.contributor.referee1.fl_str_mv Barros, Geisa Baptista
dc.contributor.referee2.fl_str_mv Fux, Blima
contributor_str_mv Pereira, Fausto Edmundo Lima
Lemos, Elenice Moreira
Barros, Geisa Baptista
Fux, Blima
dc.subject.por.fl_str_mv Toxoplasmose congênita
Citometria de fluxo
Diagnóstico
Diagnóstico sorológico
topic Toxoplasmose congênita
Citometria de fluxo
Diagnóstico
Diagnóstico sorológico
Doenças Infecciosas e Parasitárias
61
dc.subject.cnpq.fl_str_mv Doenças Infecciosas e Parasitárias
dc.subject.udc.none.fl_str_mv 61
description The routine diagnosis of pos-natal congenital toxoplasmosis is based on the detection of IgM and/or IgA anti-T. gondii antibodies in neonates sera. However, in a significant number of infected newborns these antibodies are not detected, hindering the early diagnosis of the disease. Therefore, in this study it was evaluated the performance of the detection of anti-T. gondii antibodies by flow cytometry in the sorological diagnosis of early pos-natal congenital toxoplasmosis., Eight eight sera samples of children with congenital infection by T.gondii (TOXO) and 19 sera samples of uninfected children were evaluated. The sensitivity of the test was 47.6% for IgM, 72.6% for IgA, and 75% for IgG, with 100% of specificity for all tests. As to the IgG subclasses, the sensitivity was 73.9% for IgG1, 60.2% for IgG2, and 83% for IgG3 with 100% of specificity for all tests. The sensitivity of IgG4 was superior to the aformentioned subclasses reaching 94.7%, although achieved a poor specificity of 4.6%. The IgG avidity when employed to segregate TOXO and NI groups presented a great performance, with 97% of sensitivity and 93% of specificity. It was also done a comparative analysis of the presence of IgG and IgG3 in children of TOXO and NI groups with their respective mothers. Our results showed that the children with congenital toxoplasmosis presented an average reactivity of these antibodies equivalent to their respective mothers, which suggests that they are producing antibodies in response to T.gondii infection. In contrast, uninfected children presented a lower average reactivity of those antibodies in comparison to their respective mothers. In addition, our study proposed an algorithm for the diagnosis of congenital toxoplasmosis using IgM test performed by conventional methods available in routine clinical laboratories as the initial approach followed by the search for IgG3 and IgG avidity using flow cytometry, reaching a final performance of 98% of sensitivity and 93% of specificity. Data analysis showed the applicability of flow cytometry for the detection of IgG3 antibodies and IgG anti-T. gondii avidity as a complementary tool for the early diagnosis of congenital toxoplasmosis.
publishDate 2014
dc.date.submitted.none.fl_str_mv 2014-12-04
dc.date.issued.fl_str_mv 2014-12-04
dc.date.accessioned.fl_str_mv 2015-11-16T21:07:49Z
dc.date.available.fl_str_mv 2016-06-24T06:00:07Z
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dc.format.none.fl_str_mv Text
dc.publisher.none.fl_str_mv Universidade Federal do Espírito Santo
Mestrado em Doenças Infecciosas
dc.publisher.program.fl_str_mv Programa de Pós-Graduação em Doenças Infecciosas
dc.publisher.initials.fl_str_mv UFES
dc.publisher.country.fl_str_mv BR
dc.publisher.department.fl_str_mv Centro de Ciências da Saúde
publisher.none.fl_str_mv Universidade Federal do Espírito Santo
Mestrado em Doenças Infecciosas
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