Desenvolvimento e aplicação de métodos sorológicos e moleculares para o diagnóstico da doença de Newcastle em pombos comuns (Columba livia)
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Data
2013-02-27
Autores
Oliveira, Elisabete Schirato de [UNESP]
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
Existem diversos trabalhos publicados sobre métodos de diagnóstico da Doença de Newcastle (DN) em aves comerciais da espécie Gallus gallus. Entretanto, é escassa a literatura sobre o desenvolvimento de técnicas para o diagnóstico laboratorial da infecção pelo vírus da Doença de Newcastle (VDN) em espécies de aves não-galiformes. O presente trabalho foi delineado com o objetivo de desenvolver e aplicar a técnica de bloqueio de fase líquida de ELISA com a concanavalina A (BFL-Con A-ELISA) para a detecção e quantificação de anticorpos contra o VDN em soros de pombos de vida livre e da técnica de Semi-nested-RTPCR para detecção do vírus da DN (VDN) em suabes cloacais das mesmas aves. Na comparação entre o BFL-Con A-ELISA e o HI para a detecção de anticorpos em 107 amostras de soros de pombos, foram obtidos uma correlação significativa com o teste de HI (r = 0,875), bem como valores elevados de sensibilidade (100%), especificidade (95,8%), acurácia (96,3%) e concordância (κ = 0,83). Os resultados obtidos na técnica de semi-nested-RT-PCR mostraram que nove das 101 amostras analisadas foram positivas para o VDN. Na análise comparativa dos resultados da técnica do semi-nested-RT-PCR com os obtidos nas técnicas sorológicas, foram observados índices de especificidade relativa de 93,9% e 96,8% em relação aos métodos de HI e BFL-Con A-ELISA, respectivamente; além de uma máxima sensibilidade relativa (100%), comparativamente a esses testes. Esses dados demonstram que tanto o BFL-Con A-ELISA como o semi-nested-RT-PCR desenvolvidos no presente estudo foram eficientes no diagnóstico da DN, ou detectando anticorpos anti-VDN, ou esse agente viral, respectivamente, e ambas as técnicas apresentaram um grande potencial para serem usadas com vantagens no diagnóstico da infecção pelo VDN em pombos e em outras espécies de aves não-galiformes.
A number of studies were done on diagnostic methods of Newcastle disease in commercial poultry (Gallus gallus, species). However the literature is scarce on the development of techniques for the diagnosis of Newcastle disease (ND) infection in non-galiforme and free-living birds. The present study aims to develop and apply a liquid-phase blocking ELISA with Concanavalin A (BFL-Con AELISA) for the detection and quantification of antibodies against ND virus (NDV) in sera of free-living pigeons and a semi-nested-RT-PCR technique for detection of NDV in cloacal swabs collected from these birds. The results of BFL-Con A-ELISA and HI were compared for the detection of antibodies in serum samples from 107 pigeons, and a significant correlation with the HI test was obtained (r = 0.875) as well as high values of relative sensitivity (100 %), specificity (95.8%), accuracy (96.3%) and agreement (k = 0.83). The results of semi-nested-RT-PCR showed that nine of the 101 samples were positive for the presence of NDV. By comparing the seminested- RT-PCR with serological tests, high relative specificity values of 93.9% and 96.8% were obtained with regard to HI and BLF-Con A-ELISA tests, respectively, as well as a maximum relative sensitivity (100%) was observed regarding these serological tests. These data demonstrate that the BFL-Con A-ELISA and seminested- RT-PCR developed in this study were effective in the diagnosis of NDV infection, detecting either specific antibodies, or this virus, respectively, and have a great potential to be used advantageously in the diagnostic of NDV infection in pigeons and other species of non-galliforme birds.
A number of studies were done on diagnostic methods of Newcastle disease in commercial poultry (Gallus gallus, species). However the literature is scarce on the development of techniques for the diagnosis of Newcastle disease (ND) infection in non-galiforme and free-living birds. The present study aims to develop and apply a liquid-phase blocking ELISA with Concanavalin A (BFL-Con AELISA) for the detection and quantification of antibodies against ND virus (NDV) in sera of free-living pigeons and a semi-nested-RT-PCR technique for detection of NDV in cloacal swabs collected from these birds. The results of BFL-Con A-ELISA and HI were compared for the detection of antibodies in serum samples from 107 pigeons, and a significant correlation with the HI test was obtained (r = 0.875) as well as high values of relative sensitivity (100 %), specificity (95.8%), accuracy (96.3%) and agreement (k = 0.83). The results of semi-nested-RT-PCR showed that nine of the 101 samples were positive for the presence of NDV. By comparing the seminested- RT-PCR with serological tests, high relative specificity values of 93.9% and 96.8% were obtained with regard to HI and BLF-Con A-ELISA tests, respectively, as well as a maximum relative sensitivity (100%) was observed regarding these serological tests. These data demonstrate that the BFL-Con A-ELISA and seminested- RT-PCR developed in this study were effective in the diagnosis of NDV infection, detecting either specific antibodies, or this virus, respectively, and have a great potential to be used advantageously in the diagnostic of NDV infection in pigeons and other species of non-galliforme birds.
Descrição
Palavras-chave
Teste imunoenzimático, Lectinas, Vírus da doença de Newcastle, Pombo, Newcastle disease virus
Como citar
OLIVEIRA, Elisabete Schirato de. Desenvolvimento e aplicação de métodos sorológicos e moleculares para o diagnóstico da doença de Newcastle em pombos comuns (Columba livia). 2013. xvii, 76 f. Dissertação (mestrado) - Universidade Estadual Paulista Júlio de Mesquita Filho, Faculdade de Ciências Agrárias e Veterinárias de Jaboticabal, 2013.