Desenvolvimento de um ELISA indireto com antígenos recombinantes para detecção de anticorpos contra Mycoplasma hyopneumoniae
Autor(a) principal: | |
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Data de Publicação: | 2011 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFPel - Guaiaca |
Texto Completo: | http://guaiaca.ufpel.edu.br/handle/123456789/1288 |
Resumo: | Mycoplasma hyopneumoniae is the etiological agent of swine enzootic pneumonia (EP), one of the most important respiratory diseases which affect swine worldwide. This disease is characterized by retarded development, especially in animals in the growing and finishing phase, causing significant economic losses in the herd. The presumptive diagnostic is based on clinical signs and lesions, however, laboratory tests are needed for a conclusive diagnosis of disease. Although the culture of the agent is considered the gold standard, it is not used routinely because of the slow growth of the bacterium and interference by other mycoplasmas. Therefore, there is a need for development of more sensitive and specific diagnostic methods for this disease. This study aimed the standardization and validation of an ELISA for detection of antibodies against M. hyopneumoniae. For this, six recombinant proteins of M. hyopneumoniae considered species-specific (P46, P95, P97 like, P102AB, Lppt and hypothetical protein 987) were evaluated. They were used as antigen for coating ELISA plates. Each of these proteins was confronted with three sets of pig serum. The sensitivity and specificity obtained for each protein were respectively: P95, 74.3% e 97.6%; P46, 53.7% e 97.6%; Lppt, 28.4% e 97.2%; 987, 27.1% e 96.1%; P102AB, 65.5% e 97.6%; P97 like, 72.6% e 97.2%. Using the set cut-off point, we calculated the positive predictive value, which ranged from 17.4% (for prevalence of 10%) to 99.4% (for prevalence of 90%), and negative predictive value which ranged from 34.5% to 99.5% depending on the prevalence of disease in a given area. These data show that the proteins P95, P46, P97 and P102AB like are potential targets to be use in diagnostic tests to detect antibodies against M. hyopneumoniae. |
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2014-08-20T13:32:57Z2011-06-032014-08-20T13:32:57Z2011-03-31BRUM, Clarice Brinck. Development of an ELISA with recombinant antigens to detect antibodies against Mycoplasma hyopneumoniae. 2011. 46 f. Dissertação (Mestrado em Biotecnologia) - Universidade Federal de Pelotas, Pelotas, 2011.http://guaiaca.ufpel.edu.br/handle/123456789/1288Mycoplasma hyopneumoniae is the etiological agent of swine enzootic pneumonia (EP), one of the most important respiratory diseases which affect swine worldwide. This disease is characterized by retarded development, especially in animals in the growing and finishing phase, causing significant economic losses in the herd. The presumptive diagnostic is based on clinical signs and lesions, however, laboratory tests are needed for a conclusive diagnosis of disease. Although the culture of the agent is considered the gold standard, it is not used routinely because of the slow growth of the bacterium and interference by other mycoplasmas. Therefore, there is a need for development of more sensitive and specific diagnostic methods for this disease. This study aimed the standardization and validation of an ELISA for detection of antibodies against M. hyopneumoniae. For this, six recombinant proteins of M. hyopneumoniae considered species-specific (P46, P95, P97 like, P102AB, Lppt and hypothetical protein 987) were evaluated. They were used as antigen for coating ELISA plates. Each of these proteins was confronted with three sets of pig serum. The sensitivity and specificity obtained for each protein were respectively: P95, 74.3% e 97.6%; P46, 53.7% e 97.6%; Lppt, 28.4% e 97.2%; 987, 27.1% e 96.1%; P102AB, 65.5% e 97.6%; P97 like, 72.6% e 97.2%. Using the set cut-off point, we calculated the positive predictive value, which ranged from 17.4% (for prevalence of 10%) to 99.4% (for prevalence of 90%), and negative predictive value which ranged from 34.5% to 99.5% depending on the prevalence of disease in a given area. These data show that the proteins P95, P46, P97 and P102AB like are potential targets to be use in diagnostic tests to detect antibodies against M. hyopneumoniae.O Mycoplasma hyopneumoniae é o agente etiológico da Pneumonia Enzoótica Suína (PES), uma das principais doenças respiratórias que acomete suínos no mundo. Esta doença é caracterizada por retardo no desenvolvimento, principalmente em animais em fase de crescimento e terminação, ocasionando perdas econômicas significativas no rebanho. O diagnóstico presuntivo é baseado em sinais clínicos e na presença de lesões, no entanto, testes laboratoriais são necessários para o diagnóstico conclusivo da doença. Embora o cultivo do agente seja considerado o padrão ouro, o mesmo não é usado como rotina, devido ao crescimento extremamente lento deste agente e a interferência por outros micoplasmas. Desta forma, há a necessidade do desenvolvimento de métodos de diagnósticos mais sensíveis e específicos para esta doença. Este trabalho teve como objetivo o desenvolvimento de um ELISA indireto para a detecção de anticorpos contra M. hyopneumoniae. Para isso, foram testadas individualmente seis proteínas recombinantes de M. hyopneumoniae expressas em E. coli, consideradas espécie-específica (P46, P95, P97 like, P102AB, Lppt e proteína hipotética 987), utilizadas como antígeno na sensibilização de placas de ELISA. Cada uma destas proteínas foi confrontada com 3 categorias de soros de suínos. A sensibilidade e especificidade obtida para cada uma das proteínas foram respectivamente as seguintes: P95, 74,3% e 97,6%; P46, 53,7% e 97,6%; Lppt, 28,4% e 97,2%; 987, 27,1% e 96,1%; P102AB, 65,5% e 97,6%; P97 like, 72,6% e 97,2%. Usando o ponto de corte definido, foi calculado o valor preditivo positivo que variou de 17,4% (para prevalências de 10%) a 99,4% (para prevalências de 90%), e o valor preditivo negativo que variou de 34,5% a 99,5% dependendo da prevalência da doença em uma determinada área. Estes dados mostram que proteínas como a P95, P46, P102AB e P97 like são potenciais alvos para uso em testes de diagnóstico para detecção de anticorpos contra M. hyopneumoniae.application/pdfporUniversidade Federal de PelotasPrograma de Pós-Graduação em BiotecnologiaUFPelBRBiotecnologiaMycoplasma hyopneumoniaeEnzootic pneumoniaELISADiagnosisMycoplasma hyopneumoniaeCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIADesenvolvimento de um ELISA indireto com antígenos recombinantes para detecção de anticorpos contra Mycoplasma hyopneumoniaeDevelopment of an ELISA with recombinant antigens to detect antibodies against Mycoplasma hyopneumoniaeinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesishttp://lattes.cnpq.br/9176182115504477http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4723107D9Dellagostin, Odir AntônioBrum, Clarice Brinckinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFPel - Guaiacainstname:Universidade Federal de Pelotas (UFPEL)instacron:UFPELORIGINALdissertacao_clarice_brinck_brum.pdfapplication/pdf480743http://guaiaca.ufpel.edu.br/xmlui/bitstream/123456789/1288/1/dissertacao_clarice_brinck_brum.pdff4748bb0a3a428b8f8c66eba6a7b2602MD51open accessTEXTdissertacao_clarice_brinck_brum.pdf.txtdissertacao_clarice_brinck_brum.pdf.txtExtracted Texttext/plain68606http://guaiaca.ufpel.edu.br/xmlui/bitstream/123456789/1288/2/dissertacao_clarice_brinck_brum.pdf.txtdb4adea00f9cfbbbc11788e16db22a79MD52open accessTHUMBNAILdissertacao_clarice_brinck_brum.pdf.jpgdissertacao_clarice_brinck_brum.pdf.jpgGenerated Thumbnailimage/jpeg1717http://guaiaca.ufpel.edu.br/xmlui/bitstream/123456789/1288/3/dissertacao_clarice_brinck_brum.pdf.jpgb84f6267affca3d3b3325af2d16deba6MD53open access123456789/12882019-08-23 09:22:41.877open accessoai:guaiaca.ufpel.edu.br:123456789/1288Repositório InstitucionalPUBhttp://repositorio.ufpel.edu.br/oai/requestrippel@ufpel.edu.br || repositorio@ufpel.edu.br || aline.batista@ufpel.edu.bropendoar:2019-08-23T12:22:41Repositório Institucional da UFPel - Guaiaca - Universidade Federal de Pelotas (UFPEL)false |
dc.title.por.fl_str_mv |
Desenvolvimento de um ELISA indireto com antígenos recombinantes para detecção de anticorpos contra Mycoplasma hyopneumoniae |
dc.title.alternative.eng.fl_str_mv |
Development of an ELISA with recombinant antigens to detect antibodies against Mycoplasma hyopneumoniae |
title |
Desenvolvimento de um ELISA indireto com antígenos recombinantes para detecção de anticorpos contra Mycoplasma hyopneumoniae |
spellingShingle |
Desenvolvimento de um ELISA indireto com antígenos recombinantes para detecção de anticorpos contra Mycoplasma hyopneumoniae Brum, Clarice Brinck Mycoplasma hyopneumoniae Enzootic pneumonia ELISA Diagnosis Mycoplasma hyopneumoniae CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
title_short |
Desenvolvimento de um ELISA indireto com antígenos recombinantes para detecção de anticorpos contra Mycoplasma hyopneumoniae |
title_full |
Desenvolvimento de um ELISA indireto com antígenos recombinantes para detecção de anticorpos contra Mycoplasma hyopneumoniae |
title_fullStr |
Desenvolvimento de um ELISA indireto com antígenos recombinantes para detecção de anticorpos contra Mycoplasma hyopneumoniae |
title_full_unstemmed |
Desenvolvimento de um ELISA indireto com antígenos recombinantes para detecção de anticorpos contra Mycoplasma hyopneumoniae |
title_sort |
Desenvolvimento de um ELISA indireto com antígenos recombinantes para detecção de anticorpos contra Mycoplasma hyopneumoniae |
author |
Brum, Clarice Brinck |
author_facet |
Brum, Clarice Brinck |
author_role |
author |
dc.contributor.authorID.por.fl_str_mv |
|
dc.contributor.authorLattes.por.fl_str_mv |
http://lattes.cnpq.br/9176182115504477 |
dc.contributor.advisorID.por.fl_str_mv |
|
dc.contributor.advisorLattes.por.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4723107D9 |
dc.contributor.advisor1.fl_str_mv |
Dellagostin, Odir Antônio |
dc.contributor.author.fl_str_mv |
Brum, Clarice Brinck |
contributor_str_mv |
Dellagostin, Odir Antônio |
dc.subject.eng.fl_str_mv |
Mycoplasma hyopneumoniae Enzootic pneumonia ELISA Diagnosis |
topic |
Mycoplasma hyopneumoniae Enzootic pneumonia ELISA Diagnosis Mycoplasma hyopneumoniae CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
dc.subject.por.fl_str_mv |
Mycoplasma hyopneumoniae |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
description |
Mycoplasma hyopneumoniae is the etiological agent of swine enzootic pneumonia (EP), one of the most important respiratory diseases which affect swine worldwide. This disease is characterized by retarded development, especially in animals in the growing and finishing phase, causing significant economic losses in the herd. The presumptive diagnostic is based on clinical signs and lesions, however, laboratory tests are needed for a conclusive diagnosis of disease. Although the culture of the agent is considered the gold standard, it is not used routinely because of the slow growth of the bacterium and interference by other mycoplasmas. Therefore, there is a need for development of more sensitive and specific diagnostic methods for this disease. This study aimed the standardization and validation of an ELISA for detection of antibodies against M. hyopneumoniae. For this, six recombinant proteins of M. hyopneumoniae considered species-specific (P46, P95, P97 like, P102AB, Lppt and hypothetical protein 987) were evaluated. They were used as antigen for coating ELISA plates. Each of these proteins was confronted with three sets of pig serum. The sensitivity and specificity obtained for each protein were respectively: P95, 74.3% e 97.6%; P46, 53.7% e 97.6%; Lppt, 28.4% e 97.2%; 987, 27.1% e 96.1%; P102AB, 65.5% e 97.6%; P97 like, 72.6% e 97.2%. Using the set cut-off point, we calculated the positive predictive value, which ranged from 17.4% (for prevalence of 10%) to 99.4% (for prevalence of 90%), and negative predictive value which ranged from 34.5% to 99.5% depending on the prevalence of disease in a given area. These data show that the proteins P95, P46, P97 and P102AB like are potential targets to be use in diagnostic tests to detect antibodies against M. hyopneumoniae. |
publishDate |
2011 |
dc.date.available.fl_str_mv |
2011-06-03 2014-08-20T13:32:57Z |
dc.date.issued.fl_str_mv |
2011-03-31 |
dc.date.accessioned.fl_str_mv |
2014-08-20T13:32:57Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
BRUM, Clarice Brinck. Development of an ELISA with recombinant antigens to detect antibodies against Mycoplasma hyopneumoniae. 2011. 46 f. Dissertação (Mestrado em Biotecnologia) - Universidade Federal de Pelotas, Pelotas, 2011. |
dc.identifier.uri.fl_str_mv |
http://guaiaca.ufpel.edu.br/handle/123456789/1288 |
identifier_str_mv |
BRUM, Clarice Brinck. Development of an ELISA with recombinant antigens to detect antibodies against Mycoplasma hyopneumoniae. 2011. 46 f. Dissertação (Mestrado em Biotecnologia) - Universidade Federal de Pelotas, Pelotas, 2011. |
url |
http://guaiaca.ufpel.edu.br/handle/123456789/1288 |
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por |
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por |
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info:eu-repo/semantics/openAccess |
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Universidade Federal de Pelotas |
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Programa de Pós-Graduação em Biotecnologia |
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UFPel |
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BR |
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Biotecnologia |
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Universidade Federal de Pelotas |
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