Cloning, expression and characterization of SeM protein of Streptococcus equi subsp. equi and evaluation of its use as antigen in an indirect ELISA
Strangles is an economically important horse disease caused by Streptococcus equi subsp. equi. The diagnosis can be confirmed either directly by bacterial isolation and PCR or by ELISA, which is an indirect method based on the detection of serum antibodies. The aim of this study was to clone, expres...
| Autor principal: | MORAES, Carina Martins de |
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| Outros Autores: | CONCEIÇÃO, Fabrício Rochedo, ROCHA, Andréa da Silva Ramos, SANTOS JÚNIOR, Alceu Gonçalves dos, RIBAS, Leandro dos Montes, VARGAS, Agueda Palmira Castagna de, NOGUEIRA, Carlos Eduardo Wayne, TURNES, Carlos Gil, LEITE, Fabio Pereira Leivas |
| Grau: | Artigo |
| Idioma: | eng |
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2015
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http://repositorio.ufpa.br/jspui/handle/2011/6539 |
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ir-2011-6539 |
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ir-2011-65392018-02-22T17:16:54Z Cloning, expression and characterization of SeM protein of Streptococcus equi subsp. equi and evaluation of its use as antigen in an indirect ELISA Clonagem, expressão e caracterização de proteína SeM de Streptococcus equi subsp. equi e avaliação de sua utilização como antígeno em um ELISA indireto MORAES, Carina Martins de CONCEIÇÃO, Fabrício Rochedo ROCHA, Andréa da Silva Ramos SANTOS JÚNIOR, Alceu Gonçalves dos RIBAS, Leandro dos Montes VARGAS, Agueda Palmira Castagna de NOGUEIRA, Carlos Eduardo Wayne TURNES, Carlos Gil LEITE, Fabio Pereira Leivas Medicina veterinária Equino Adenite equina Streptococcus equi SeM Ensaio de imunoadsorção enzimática Strangles is an economically important horse disease caused by Streptococcus equi subsp. equi. The diagnosis can be confirmed either directly by bacterial isolation and PCR or by ELISA, which is an indirect method based on the detection of serum antibodies. The aim of this study was to clone, express and characterize the SeM protein of Streptococcus equi subsp. equi, evaluate its use as antigen in indirect ELISA and determine its performance to distinguish sera of negative, vaccinated and positive animals. This was initially performed by cloning the gene encoding the SeM protein and its expression in Escherichia coli. Subsequently, the protein produced was characterized and used as antigen in ELISA. Serum samples for evaluation were taken from 40 negative foals, 46 horses vaccinated with a commercial vaccine against strangles and 46 horses diagnosed with the disease. The test showed high specificity and sensitivity, allowing discrimination between negative and positive, positive and vaccinated animals, and vaccinated animals and negative sera. Thus, it was concluded that the protein produced rSeM, which can be used as antigen for disease diagnosis, and the described ELISA might be helpful to evaluate the immune status of the herd. A adenite equina é uma enfermidade economicamente importante de equinos, causada por Streptococcus equi subsp. equi. Seu diagnóstico pode ser confirmado de forma direta, por meio de isolamento bacteriano e de PCR, ou de forma indireta, por meio de ELISA, método baseado na detecção de anticorpos séricos. O objetivo deste estudo foi clonar, expressar e caracterizar a proteína SeM de Streptococcus equi subsp. equi, avaliar sua utilização como antígeno em um ELISA indireto e determinar a capacidade do teste de distinguir soros de animais negativos, vacinados e positivos. Para tal, foi inicialmente realizada a clonagem do gene que codifica para a proteína SeM e sua expressão em Escherichia coli. Posteriormente, a proteína produzida foi caracterizada e utilizada como antígeno em um teste de ELISA indireto. Para avaliação do teste, foram utilizadas amostras de soro de 40 potros negativos, de 46 equinos vacinados com uma vacina comercial contra adenite equina e de 46 equinos com diagnóstico da doença. O teste demonstrou alta sensibilidade e especificidade, permitindo discriminar entre soros negativos e positivos, positivos e de animais vacinados, e negativos e de animais vacinados. Assim, conclui-se que a proteína rSeM produzida pode ser usada como antígeno para o diagnóstico da enfermidade e que o ELISA descrito pode ser útil para avaliar o estado imunológico do rebanho. 2015-04-14T13:51:34Z 2015-04-14T13:51:34Z 2014-08 Artigo de Periódico MORAES, C.M. et al. Cloning, expression and characterization of SeM protein of Streptococcus equi subsp. equi and evaluation of its use as antigen in an indirect ELISA. Arquivo Brasileiro de Medicina Veterinária e Zootecnia, Belo Horizonte, v. 66, n. 4, p. 1015-1022, ago. 2014. Disponível em: <http://www.scielo.br/pdf/abmvz/v66n4/0102-0935-abmvz-66-04-01015.pdf>. Acesso em: 13 abr. 2015. <http://dx.doi.org/10.1590/1678-6034>. 0102-0935 http://repositorio.ufpa.br/jspui/handle/2011/6539 eng Acesso Aberto application/pdf |
| institution |
Repositório Institucional - Universidade Federal do Pará |
| collection |
RI-UFPA |
| language |
eng |
| topic |
Medicina veterinária Equino Adenite equina Streptococcus equi SeM Ensaio de imunoadsorção enzimática |
| spellingShingle |
Medicina veterinária Equino Adenite equina Streptococcus equi SeM Ensaio de imunoadsorção enzimática MORAES, Carina Martins de Cloning, expression and characterization of SeM protein of Streptococcus equi subsp. equi and evaluation of its use as antigen in an indirect ELISA |
| topic_facet |
Medicina veterinária Equino Adenite equina Streptococcus equi SeM Ensaio de imunoadsorção enzimática |
| description |
Strangles is an economically important horse disease caused by Streptococcus equi subsp. equi. The diagnosis can be confirmed either directly by bacterial isolation and PCR or by ELISA, which is an indirect method based on the detection of serum antibodies. The aim of this study was to clone, express and characterize the SeM protein of Streptococcus equi subsp. equi, evaluate its use as antigen in indirect ELISA and determine its performance to distinguish sera of negative, vaccinated and positive animals. This was initially performed by cloning the gene encoding the SeM protein and its expression in Escherichia coli. Subsequently, the protein produced was characterized and used as antigen in ELISA. Serum samples for evaluation were taken from 40 negative foals, 46 horses vaccinated with a commercial vaccine against strangles and 46 horses diagnosed with the disease. The test showed high specificity and sensitivity, allowing discrimination between negative and positive, positive and vaccinated animals, and vaccinated animals and negative sera. Thus, it was concluded that the protein produced rSeM, which can be used as antigen for disease diagnosis, and the described ELISA might be helpful to evaluate the immune status of the herd. |
| format |
Artigo |
| author |
MORAES, Carina Martins de |
| author2 |
CONCEIÇÃO, Fabrício Rochedo ROCHA, Andréa da Silva Ramos SANTOS JÚNIOR, Alceu Gonçalves dos RIBAS, Leandro dos Montes VARGAS, Agueda Palmira Castagna de NOGUEIRA, Carlos Eduardo Wayne TURNES, Carlos Gil LEITE, Fabio Pereira Leivas |
| author2Str |
CONCEIÇÃO, Fabrício Rochedo ROCHA, Andréa da Silva Ramos SANTOS JÚNIOR, Alceu Gonçalves dos RIBAS, Leandro dos Montes VARGAS, Agueda Palmira Castagna de NOGUEIRA, Carlos Eduardo Wayne TURNES, Carlos Gil LEITE, Fabio Pereira Leivas |
| title |
Cloning, expression and characterization of SeM protein of Streptococcus equi subsp. equi and evaluation of its use as antigen in an indirect ELISA |
| title_short |
Cloning, expression and characterization of SeM protein of Streptococcus equi subsp. equi and evaluation of its use as antigen in an indirect ELISA |
| title_full |
Cloning, expression and characterization of SeM protein of Streptococcus equi subsp. equi and evaluation of its use as antigen in an indirect ELISA |
| title_fullStr |
Cloning, expression and characterization of SeM protein of Streptococcus equi subsp. equi and evaluation of its use as antigen in an indirect ELISA |
| title_full_unstemmed |
Cloning, expression and characterization of SeM protein of Streptococcus equi subsp. equi and evaluation of its use as antigen in an indirect ELISA |
| title_sort |
cloning, expression and characterization of sem protein of streptococcus equi subsp. equi and evaluation of its use as antigen in an indirect elisa |
| publishDate |
2015 |
| url |
http://repositorio.ufpa.br/jspui/handle/2011/6539 |
| _version_ |
1832604907115380736 |
| score |
11.755432 |