Estudo do Circovírus suíno - 2 (PCV2) pelas técnicas de ELISA indireto por captura de anticorpo e PCR em tempo real em suínos

Detalhes bibliográficos
Autor(a) principal: BARBOSA JUNIOR, Sebastião André
Data de Publicação: 2016
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Biblioteca Digital de Teses e Dissertações da UFRPE
Texto Completo: http://www.tede2.ufrpe.br:8080/tede2/handle/tede2/7151
Resumo: Porcine circovirus (PCV) belongs to the Circoviridae family and Circovirus genus. The vírus can be subdivided into porcine circovirus-1 (PCV1) and porcine circovirus-2 (PCV2). PCV1 is non-pathogenic, where as PCV2 is one of the most important etiological agents in the pig industry today, and the cause of several diseases. The complexity of PCV2 caused by disturbances has led to a demand for laboratory techniques and a wider range of biological samples that can facilitate diagnosis and herd health monitoring. Research studies of the antibodies against PCV2 have prefered the ELISA technique to other serological methods because of its simplicity, high sensitivity and specificity. Among the molecular tools, the quantitative real-time polymerase chain reaction (qPCR) stands out since it makes it possible to measure the viral load of the sample. The oral fluid (FO) is a biological sample that has been increasingly used in pig farming as it reduces stress and improves animal welfare. Although PCV2 has been recently detected in Brazil, the North-East region has still not carried out many studies on the dynamics of the respective viruses. In view of this, the, aim of this research is to study PCV2 by means of indirect ELISA with the capture antibody and qPCR in pigs through serum and FO, respectively. The samples were obtained from the Municipal Slaughterhouse in the town of Paulista, located in the metropolitan district of Recife, Pernambuco State. Serum samples were collected during the bleeding process, and a total of 74 samples were obtained. These were assessed by means of the ELISA technique with the capture antibody. The FO samples were obtained from pigs that were housed in stalls to rest. A total of 60 samples were collected, and these were subjected to qPCR testing. The serum samples that were analyzed, had measurements in the ELISA index as follows: very low - 14 (18.92%), low - 13 (17.57%), moderate - 11 (14.86%) and high - 36 (48.65%). The FO samples showed the DNA of the virus as 13.34% (8/60). 75% (6/8) of these had a very low viral load of 4 (log) / ml of the sample and a low viral load 25% (2/8) 4 to 5 (log) / ml sample. The occurrence of antibodies and the presence of PCV2 in the FO samples suggest that the virus is spreading in the herds of Pernambuco State. This study provides a strong argument for the application of ELISA for capture antibody in serological tests and using pids to obtain biological FO samples in quantitative tests.
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spelling BARBOSA, Clara NilceFREITAS, Antônio Carlos deFREITAS, Aderaldo Alexandrino dehttp://lattes.cnpq.br/2696046779149842BARBOSA JUNIOR, Sebastião André2018-02-27T14:17:28Z2016-02-26BARBOSA JÚNIOR, Sebastião André. Estudo do Circovírus suíno - 2 (PCV2) pelas técnicas de ELISA indireto por captura de anticorpo e PCR em tempo real em suínos. 2016. 52 f. Dissertação (Programa de Pós-Graduação em Ciência Veterinária) - Universidade Federal Rural de Pernambuco, Recife.http://www.tede2.ufrpe.br:8080/tede2/handle/tede2/7151Porcine circovirus (PCV) belongs to the Circoviridae family and Circovirus genus. The vírus can be subdivided into porcine circovirus-1 (PCV1) and porcine circovirus-2 (PCV2). PCV1 is non-pathogenic, where as PCV2 is one of the most important etiological agents in the pig industry today, and the cause of several diseases. The complexity of PCV2 caused by disturbances has led to a demand for laboratory techniques and a wider range of biological samples that can facilitate diagnosis and herd health monitoring. Research studies of the antibodies against PCV2 have prefered the ELISA technique to other serological methods because of its simplicity, high sensitivity and specificity. Among the molecular tools, the quantitative real-time polymerase chain reaction (qPCR) stands out since it makes it possible to measure the viral load of the sample. The oral fluid (FO) is a biological sample that has been increasingly used in pig farming as it reduces stress and improves animal welfare. Although PCV2 has been recently detected in Brazil, the North-East region has still not carried out many studies on the dynamics of the respective viruses. In view of this, the, aim of this research is to study PCV2 by means of indirect ELISA with the capture antibody and qPCR in pigs through serum and FO, respectively. The samples were obtained from the Municipal Slaughterhouse in the town of Paulista, located in the metropolitan district of Recife, Pernambuco State. Serum samples were collected during the bleeding process, and a total of 74 samples were obtained. These were assessed by means of the ELISA technique with the capture antibody. The FO samples were obtained from pigs that were housed in stalls to rest. A total of 60 samples were collected, and these were subjected to qPCR testing. The serum samples that were analyzed, had measurements in the ELISA index as follows: very low - 14 (18.92%), low - 13 (17.57%), moderate - 11 (14.86%) and high - 36 (48.65%). The FO samples showed the DNA of the virus as 13.34% (8/60). 75% (6/8) of these had a very low viral load of 4 (log) / ml of the sample and a low viral load 25% (2/8) 4 to 5 (log) / ml sample. The occurrence of antibodies and the presence of PCV2 in the FO samples suggest that the virus is spreading in the herds of Pernambuco State. This study provides a strong argument for the application of ELISA for capture antibody in serological tests and using pids to obtain biological FO samples in quantitative tests.O Circovírus suíno (PCV) pertence à família Circoviridae e ao genêro Circovírus. O vírus subdivide-se em: Circovírus suíno-1 (PCV1) e Circovírus suíno-2 (PCV2). O PCV1 não é patogênico, ao contrário do PCV2, que é um dos mais importantes agentes etiológicos da suinocultura moderna, estando associado a várias doenças. Essa complexidade de distúrbios causados pelo PCV2 gera uma demanda por técnicas laboratoriais e mais opções de amostras biológicas que venham a facilitar o diagnóstico e monitoramento sanitário dos rebanhos. A técnica do ELISA vem destacando-se na investigação de anticorpos contra o PCV2, diante de outras técnicas sorológicas, em razão de sua simplicidade, alta sensibilidade e especificidade. Dentre as ferramentas moleculares, a Reação em Cadeia pela Polimerase quantitativa em tempo real (qPCR) destaca-se principalmente por mensurar a carga viral da amostra. O fluido oral (FO) é uma amostra biológica que vem sendo cada vez mais utilizada na suinocultura por diminuir o estresse e favorecer o bem-estar animal. O PCV2 foi identificado recentemente no Brasil, e a região Nordeste ainda apresenta poucos estudos sobre a dinâmica do respectivo vírus. Baseado nesses aspectos, objetivou-se estudar o PCV2 pelas técnicas de ELISA indireto com anticorpo de captura e de qPCR em suínos, por meio de amostras de soro e FO, respectivamente. As amostras foram oriundas do Abatedouro Municipal da cidade de Paulista, localizado na Região Metropolitana do Recife, estado de Pernambuco. As amostras de soro foram coletadas durante o processo da sangria, no qual foi obtido um total de 74 amostras. Essas foram avaliadas pela técnica do ELISA indireto com anticorpo de captura. As amostras de FO foram obtidas de suínos que estavam alojados nas baias de descanso. Foi coletado um total de 60 amostras, as quais foram submetidas ao teste do qPCR. Das amostras de soro analisadas, tiveram o índice do ELISA considerado baixíssimo 14 (18,92%), baixo 13 (17,57%), moderado 11 (14,86%) e alto 36 (48,65%). Das amostras de FO, apresentaram o DNA do vírus 13,34% (8/60). Dessas, 75% (6/8) apresentaram uma carga viral muito baixa, até 4 (log)/mL da amostra, e uma carga viral baixa 25% (2/8), de 4 a 5 (log)/mL da amostra. A ocorrência de anticorpos e a presença do PCV2 nas amostras de FO sugerem que o vírus está circulante em rebanhos do estado de Pernambuco. Este estudo traz argumentos para a aplicação do ELISA indireto por anticorpo de captura em pesquisas sorológicas e utilização do FO de suíno como amostra biológica em testes quantitativos.Submitted by Mario BC (mario@bc.ufrpe.br) on 2018-02-27T14:16:51Z No. of bitstreams: 1 Sebastiao Andre Barbosa Junior.pdf: 749859 bytes, checksum: e483888ecfa0c1a9815a1e63aaede716 (MD5)Made available in DSpace on 2018-02-27T14:17:28Z (GMT). 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dc.title.por.fl_str_mv Estudo do Circovírus suíno - 2 (PCV2) pelas técnicas de ELISA indireto por captura de anticorpo e PCR em tempo real em suínos
title Estudo do Circovírus suíno - 2 (PCV2) pelas técnicas de ELISA indireto por captura de anticorpo e PCR em tempo real em suínos
spellingShingle Estudo do Circovírus suíno - 2 (PCV2) pelas técnicas de ELISA indireto por captura de anticorpo e PCR em tempo real em suínos
BARBOSA JUNIOR, Sebastião André
Circovírus suíno
Carga viral
Diagnóstico
Teste ELISA
Suíno
CIENCIAS AGRARIAS::MEDICINA VETERINARIA
title_short Estudo do Circovírus suíno - 2 (PCV2) pelas técnicas de ELISA indireto por captura de anticorpo e PCR em tempo real em suínos
title_full Estudo do Circovírus suíno - 2 (PCV2) pelas técnicas de ELISA indireto por captura de anticorpo e PCR em tempo real em suínos
title_fullStr Estudo do Circovírus suíno - 2 (PCV2) pelas técnicas de ELISA indireto por captura de anticorpo e PCR em tempo real em suínos
title_full_unstemmed Estudo do Circovírus suíno - 2 (PCV2) pelas técnicas de ELISA indireto por captura de anticorpo e PCR em tempo real em suínos
title_sort Estudo do Circovírus suíno - 2 (PCV2) pelas técnicas de ELISA indireto por captura de anticorpo e PCR em tempo real em suínos
author BARBOSA JUNIOR, Sebastião André
author_facet BARBOSA JUNIOR, Sebastião André
author_role author
dc.contributor.advisor1.fl_str_mv BARBOSA, Clara Nilce
dc.contributor.advisor-co1.fl_str_mv FREITAS, Antônio Carlos de
dc.contributor.referee1.fl_str_mv FREITAS, Aderaldo Alexandrino de
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/2696046779149842
dc.contributor.author.fl_str_mv BARBOSA JUNIOR, Sebastião André
contributor_str_mv BARBOSA, Clara Nilce
FREITAS, Antônio Carlos de
FREITAS, Aderaldo Alexandrino de
dc.subject.por.fl_str_mv Circovírus suíno
Carga viral
Diagnóstico
Teste ELISA
Suíno
topic Circovírus suíno
Carga viral
Diagnóstico
Teste ELISA
Suíno
CIENCIAS AGRARIAS::MEDICINA VETERINARIA
dc.subject.cnpq.fl_str_mv CIENCIAS AGRARIAS::MEDICINA VETERINARIA
description Porcine circovirus (PCV) belongs to the Circoviridae family and Circovirus genus. The vírus can be subdivided into porcine circovirus-1 (PCV1) and porcine circovirus-2 (PCV2). PCV1 is non-pathogenic, where as PCV2 is one of the most important etiological agents in the pig industry today, and the cause of several diseases. The complexity of PCV2 caused by disturbances has led to a demand for laboratory techniques and a wider range of biological samples that can facilitate diagnosis and herd health monitoring. Research studies of the antibodies against PCV2 have prefered the ELISA technique to other serological methods because of its simplicity, high sensitivity and specificity. Among the molecular tools, the quantitative real-time polymerase chain reaction (qPCR) stands out since it makes it possible to measure the viral load of the sample. The oral fluid (FO) is a biological sample that has been increasingly used in pig farming as it reduces stress and improves animal welfare. Although PCV2 has been recently detected in Brazil, the North-East region has still not carried out many studies on the dynamics of the respective viruses. In view of this, the, aim of this research is to study PCV2 by means of indirect ELISA with the capture antibody and qPCR in pigs through serum and FO, respectively. The samples were obtained from the Municipal Slaughterhouse in the town of Paulista, located in the metropolitan district of Recife, Pernambuco State. Serum samples were collected during the bleeding process, and a total of 74 samples were obtained. These were assessed by means of the ELISA technique with the capture antibody. The FO samples were obtained from pigs that were housed in stalls to rest. A total of 60 samples were collected, and these were subjected to qPCR testing. The serum samples that were analyzed, had measurements in the ELISA index as follows: very low - 14 (18.92%), low - 13 (17.57%), moderate - 11 (14.86%) and high - 36 (48.65%). The FO samples showed the DNA of the virus as 13.34% (8/60). 75% (6/8) of these had a very low viral load of 4 (log) / ml of the sample and a low viral load 25% (2/8) 4 to 5 (log) / ml sample. The occurrence of antibodies and the presence of PCV2 in the FO samples suggest that the virus is spreading in the herds of Pernambuco State. This study provides a strong argument for the application of ELISA for capture antibody in serological tests and using pids to obtain biological FO samples in quantitative tests.
publishDate 2016
dc.date.issued.fl_str_mv 2016-02-26
dc.date.accessioned.fl_str_mv 2018-02-27T14:17:28Z
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dc.identifier.citation.fl_str_mv BARBOSA JÚNIOR, Sebastião André. Estudo do Circovírus suíno - 2 (PCV2) pelas técnicas de ELISA indireto por captura de anticorpo e PCR em tempo real em suínos. 2016. 52 f. Dissertação (Programa de Pós-Graduação em Ciência Veterinária) - Universidade Federal Rural de Pernambuco, Recife.
dc.identifier.uri.fl_str_mv http://www.tede2.ufrpe.br:8080/tede2/handle/tede2/7151
identifier_str_mv BARBOSA JÚNIOR, Sebastião André. Estudo do Circovírus suíno - 2 (PCV2) pelas técnicas de ELISA indireto por captura de anticorpo e PCR em tempo real em suínos. 2016. 52 f. Dissertação (Programa de Pós-Graduação em Ciência Veterinária) - Universidade Federal Rural de Pernambuco, Recife.
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