Validation of a Real Time PCR for Classical Swine Fever Diagnosis
The viral disease classical swine fever (CSF), caused by a Pestivirus, is one of the major causes of economic losses for pig farming. The aim of this work was to validate a RT-qPCR using Taqman for detection of CSF in swine tissues. The parameters for the validation followed the specifications of th...
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| Format: | Article |
| Language: | English |
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Wiley
2014-01-01
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| Series: | Veterinary Medicine International |
| Online Access: | http://dx.doi.org/10.1155/2014/171235 |
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| author | Natanael Lamas Dias Antônio Augusto Fonseca Júnior Anapolino Macedo Oliveira Érica Bravo Sales Bruna Rios Coelho Alves Fernanda Alves Dorella Marcelo Fernandes Camargos |
| author_facet | Natanael Lamas Dias Antônio Augusto Fonseca Júnior Anapolino Macedo Oliveira Érica Bravo Sales Bruna Rios Coelho Alves Fernanda Alves Dorella Marcelo Fernandes Camargos |
| author_sort | Natanael Lamas Dias |
| collection | DOAJ |
| description | The viral disease classical swine fever (CSF), caused by a Pestivirus, is one of the major causes of economic losses for pig farming. The aim of this work was to validate a RT-qPCR using Taqman for detection of CSF in swine tissues. The parameters for the validation followed the specifications of the Manual of Diagnostic Tests and Vaccines for Terrestrial Animals of the World Organization for Animal Health (OIE) and the guide ABNT NBR ISO/IEC 17025:2005. The analysis of the 5′NTR region of CSF virus was performed in 145 samples from 29 infected pigs and in 240 samples from 80 pigs originated in the Brazilian CSF-free zone. The tissues tested were spleen, kidney, blood, tonsils, and lymph nodes. Sequencing of the positive samples for 5′NTR region was performed to evaluate the specificity of the RT-qPCR. Tests performed for the RT-qPCR validation demonstrated that the PCR assay was efficient in detecting RNA from CSF virus in all materials from different tissues of infected animals. Furthermore, RNA from CSF virus was not detected in samples of swine originated from the Brazilian CSF-free zone. Hence, it is concluded that RT-qPCR can be used as a complementary diagnostic for CSF. |
| format | Article |
| id | doaj-art-d87d95f418894f3ea15c4913bb1d1d7d |
| institution | Kabale University |
| issn | 2090-8113 2042-0048 |
| language | English |
| publishDate | 2014-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Veterinary Medicine International |
| spelling | doaj-art-d87d95f418894f3ea15c4913bb1d1d7d2025-02-03T07:24:05ZengWileyVeterinary Medicine International2090-81132042-00482014-01-01201410.1155/2014/171235171235Validation of a Real Time PCR for Classical Swine Fever DiagnosisNatanael Lamas Dias0Antônio Augusto Fonseca Júnior1Anapolino Macedo Oliveira2Érica Bravo Sales3Bruna Rios Coelho Alves4Fernanda Alves Dorella5Marcelo Fernandes Camargos6Laboratório de Biologia Molecular e Laboratório de Diagnóstico de Doenás Virais, Laboratório Nacional Agropecuário-Minas Gerais (LANAGRO-MG), Ministério da Agricultura, Pecuária e Abastecimento, Avenida Rômulo Joviano s/n, Fazenda Modelo, 33600-000 Pedro Leopoldo, MG, BrazilLaboratório de Biologia Molecular e Laboratório de Diagnóstico de Doenás Virais, Laboratório Nacional Agropecuário-Minas Gerais (LANAGRO-MG), Ministério da Agricultura, Pecuária e Abastecimento, Avenida Rômulo Joviano s/n, Fazenda Modelo, 33600-000 Pedro Leopoldo, MG, BrazilLaboratório de Biologia Molecular e Laboratório de Diagnóstico de Doenás Virais, Laboratório Nacional Agropecuário-Minas Gerais (LANAGRO-MG), Ministério da Agricultura, Pecuária e Abastecimento, Avenida Rômulo Joviano s/n, Fazenda Modelo, 33600-000 Pedro Leopoldo, MG, BrazilLaboratório de Biologia Molecular e Laboratório de Diagnóstico de Doenás Virais, Laboratório Nacional Agropecuário-Minas Gerais (LANAGRO-MG), Ministério da Agricultura, Pecuária e Abastecimento, Avenida Rômulo Joviano s/n, Fazenda Modelo, 33600-000 Pedro Leopoldo, MG, BrazilEmbrapa Dairy Cattle, Rua Eugênio do Nascimento 610, 36038-330 Juiz de Fora, MG, BrazilLaboratório de Biologia Molecular e Laboratório de Diagnóstico de Doenás Virais, Laboratório Nacional Agropecuário-Minas Gerais (LANAGRO-MG), Ministério da Agricultura, Pecuária e Abastecimento, Avenida Rômulo Joviano s/n, Fazenda Modelo, 33600-000 Pedro Leopoldo, MG, BrazilLaboratório de Biologia Molecular e Laboratório de Diagnóstico de Doenás Virais, Laboratório Nacional Agropecuário-Minas Gerais (LANAGRO-MG), Ministério da Agricultura, Pecuária e Abastecimento, Avenida Rômulo Joviano s/n, Fazenda Modelo, 33600-000 Pedro Leopoldo, MG, BrazilThe viral disease classical swine fever (CSF), caused by a Pestivirus, is one of the major causes of economic losses for pig farming. The aim of this work was to validate a RT-qPCR using Taqman for detection of CSF in swine tissues. The parameters for the validation followed the specifications of the Manual of Diagnostic Tests and Vaccines for Terrestrial Animals of the World Organization for Animal Health (OIE) and the guide ABNT NBR ISO/IEC 17025:2005. The analysis of the 5′NTR region of CSF virus was performed in 145 samples from 29 infected pigs and in 240 samples from 80 pigs originated in the Brazilian CSF-free zone. The tissues tested were spleen, kidney, blood, tonsils, and lymph nodes. Sequencing of the positive samples for 5′NTR region was performed to evaluate the specificity of the RT-qPCR. Tests performed for the RT-qPCR validation demonstrated that the PCR assay was efficient in detecting RNA from CSF virus in all materials from different tissues of infected animals. Furthermore, RNA from CSF virus was not detected in samples of swine originated from the Brazilian CSF-free zone. Hence, it is concluded that RT-qPCR can be used as a complementary diagnostic for CSF.http://dx.doi.org/10.1155/2014/171235 |
| spellingShingle | Natanael Lamas Dias Antônio Augusto Fonseca Júnior Anapolino Macedo Oliveira Érica Bravo Sales Bruna Rios Coelho Alves Fernanda Alves Dorella Marcelo Fernandes Camargos Validation of a Real Time PCR for Classical Swine Fever Diagnosis Veterinary Medicine International |
| title | Validation of a Real Time PCR for Classical Swine Fever Diagnosis |
| title_full | Validation of a Real Time PCR for Classical Swine Fever Diagnosis |
| title_fullStr | Validation of a Real Time PCR for Classical Swine Fever Diagnosis |
| title_full_unstemmed | Validation of a Real Time PCR for Classical Swine Fever Diagnosis |
| title_short | Validation of a Real Time PCR for Classical Swine Fever Diagnosis |
| title_sort | validation of a real time pcr for classical swine fever diagnosis |
| url | http://dx.doi.org/10.1155/2014/171235 |
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