Improvement of bovine pestiviral diagnosis by the development of a cost-effective method for detecting viral RNA in fresh specimens and samples spotted in filter papers
Bovine pestiviruses are the causative agents of bovine viral diarrhea, a disease thatcauses severe economic losses in cattle. The aim of this study was to improve their diagnosisby developing a RT-qPCR to detect bovine pestiviruses A, B and H; and to set up a protocolfor collecting, shipping and pre...
| Autores principales: | , , , , |
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| Formato: | info:ar-repo/semantics/artículo |
| Lenguaje: | Inglés |
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Elsevier
2023
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| Materias: | |
| Acceso en línea: | http://hdl.handle.net/20.500.12123/14582 https://www.sciencedirect.com/science/article/pii/S0325754122001043 https://doi.org/10.1016/j.ram.2022.10.002 |
| _version_ | 1855037142915350528 |
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| author | Favaro, Paula Melisa Molineri, Ana Ines Dus Santos, Maria Jose Calvinho, Fernando Luis Pecora, Andrea |
| author_browse | Calvinho, Fernando Luis Dus Santos, Maria Jose Favaro, Paula Melisa Molineri, Ana Ines Pecora, Andrea |
| author_facet | Favaro, Paula Melisa Molineri, Ana Ines Dus Santos, Maria Jose Calvinho, Fernando Luis Pecora, Andrea |
| author_sort | Favaro, Paula Melisa |
| collection | INTA Digital |
| description | Bovine pestiviruses are the causative agents of bovine viral diarrhea, a disease thatcauses severe economic losses in cattle. The aim of this study was to improve their diagnosisby developing a RT-qPCR to detect bovine pestiviruses A, B and H; and to set up a protocolfor collecting, shipping and preserving bovine pestiviral RNA on filter papers. The developedRT-qPCR showed high sensitivity in detecting these viruses in different matrices: viral stocks,semen and serum samples. With regard to the possibility of using the technique to test serumpools, it was possible to identify a positive serum sample within a pool containing 30 sera.In addition to evaluating the qPCR from fresh samples, the use of filter papers to sow bovinesamples was analyzed. The sampling method on two different filter papers using bovine blooddrops was a useful alternative for diagnostic purposes and allowed to preserve pestiviral RNAfor up to 12 months under refrigeration. |
| format | info:ar-repo/semantics/artículo |
| id | INTA14582 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2023 |
| publishDateRange | 2023 |
| publishDateSort | 2023 |
| publisher | Elsevier |
| publisherStr | Elsevier |
| record_format | dspace |
| spelling | INTA145822023-04-26T16:49:48Z Improvement of bovine pestiviral diagnosis by the development of a cost-effective method for detecting viral RNA in fresh specimens and samples spotted in filter papers Favaro, Paula Melisa Molineri, Ana Ines Dus Santos, Maria Jose Calvinho, Fernando Luis Pecora, Andrea Diagnosis Bovine Diarrhoea Pestivirus Animal Diseases Diagnóstico Pestivirus de la Diarrea Bovina Enfermedades de los Animales Bovine Pestiviruses Filter Papers RT-qPCR Bovine Viral Diaorrhea Pestivirus de Bovinos Papeles de Filtro Diarrea Viral Bovina Bovine pestiviruses are the causative agents of bovine viral diarrhea, a disease thatcauses severe economic losses in cattle. The aim of this study was to improve their diagnosisby developing a RT-qPCR to detect bovine pestiviruses A, B and H; and to set up a protocolfor collecting, shipping and preserving bovine pestiviral RNA on filter papers. The developedRT-qPCR showed high sensitivity in detecting these viruses in different matrices: viral stocks,semen and serum samples. With regard to the possibility of using the technique to test serumpools, it was possible to identify a positive serum sample within a pool containing 30 sera.In addition to evaluating the qPCR from fresh samples, the use of filter papers to sow bovinesamples was analyzed. The sampling method on two different filter papers using bovine blooddrops was a useful alternative for diagnostic purposes and allowed to preserve pestiviral RNAfor up to 12 months under refrigeration. Los Pestivirus bovinos son los agentes causales de la diarrea viral bovina, una enfermedad que genera importantes pérdidas económicas en el ganado vacuno. El objetivo de este trabajo fue mejorar su diagnóstico mediante el desarrollo de una RT-qPCR para detectar los Pestivirus bovinos A, B y H y diseñar un protocolo de recolección, envío y conservación de ARN viral en papeles de filtro. La RT-qPCR desarrollada demostró alta sensibilidad en la detección de estos virus en diferentes matrices: stock viral, suero y semen. Respecto de la posibilidad de usar la técnica para testear pools de suero, fue posible identificar un suero positivo dentro de un pool compuesto por 30 sueros. Además de evaluar la qPCR en muestras frescas, se analizó el uso de papeles de filtro para sembrar muestras de bovinos. La metodología de toma de muestras en dos tipos de papeles de filtro usando gotas de sangre fue una alternativa útil para el diagnóstico y permitió conservar ARN viral por hasta 12 meses a temperaturas de refrigeración. EEA Rafaela Fil: Favaro, Paula M. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias; Argentina Fil: Molineri, Ana Ines. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina Fil: Molineri, Ana Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina Fil: Dus Santos, Maria J. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas (IVIT); Argentina Fil: Dus Santos, Maria J. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Calvinho, Luis. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina Fil: Calvinho, Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina Fil: Calvinho, Luis. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias; Argentina Fil: Pecora, Andrea. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas (IVIT); Argentina Fil: Pecora, Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina 2023-04-26T16:29:46Z 2023-04-26T16:29:46Z 2023-01 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/acceptedVersion http://hdl.handle.net/20.500.12123/14582 https://www.sciencedirect.com/science/article/pii/S0325754122001043 P.M. Favaro, A. Molineri, M.J. Dus Santos et al., Improvement of bovine pestiviral diagnosis by the development of a cost-effective method for detecting viral RNA in fresh specimens and samples spotted in filter papers, Revista Argentina de Microbiología, https://doi.org/10.1016/j.ram.2022.10.002 0325-7541 https://doi.org/10.1016/j.ram.2022.10.002 eng info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf Elsevier Revista Argentina de Microbiología : 1-9 (Available online 17 January 2023) |
| spellingShingle | Diagnosis Bovine Diarrhoea Pestivirus Animal Diseases Diagnóstico Pestivirus de la Diarrea Bovina Enfermedades de los Animales Bovine Pestiviruses Filter Papers RT-qPCR Bovine Viral Diaorrhea Pestivirus de Bovinos Papeles de Filtro Diarrea Viral Bovina Favaro, Paula Melisa Molineri, Ana Ines Dus Santos, Maria Jose Calvinho, Fernando Luis Pecora, Andrea Improvement of bovine pestiviral diagnosis by the development of a cost-effective method for detecting viral RNA in fresh specimens and samples spotted in filter papers |
| title | Improvement of bovine pestiviral diagnosis by the development of a cost-effective method for detecting viral RNA in fresh specimens and samples spotted in filter papers |
| title_full | Improvement of bovine pestiviral diagnosis by the development of a cost-effective method for detecting viral RNA in fresh specimens and samples spotted in filter papers |
| title_fullStr | Improvement of bovine pestiviral diagnosis by the development of a cost-effective method for detecting viral RNA in fresh specimens and samples spotted in filter papers |
| title_full_unstemmed | Improvement of bovine pestiviral diagnosis by the development of a cost-effective method for detecting viral RNA in fresh specimens and samples spotted in filter papers |
| title_short | Improvement of bovine pestiviral diagnosis by the development of a cost-effective method for detecting viral RNA in fresh specimens and samples spotted in filter papers |
| title_sort | improvement of bovine pestiviral diagnosis by the development of a cost effective method for detecting viral rna in fresh specimens and samples spotted in filter papers |
| topic | Diagnosis Bovine Diarrhoea Pestivirus Animal Diseases Diagnóstico Pestivirus de la Diarrea Bovina Enfermedades de los Animales Bovine Pestiviruses Filter Papers RT-qPCR Bovine Viral Diaorrhea Pestivirus de Bovinos Papeles de Filtro Diarrea Viral Bovina |
| url | http://hdl.handle.net/20.500.12123/14582 https://www.sciencedirect.com/science/article/pii/S0325754122001043 https://doi.org/10.1016/j.ram.2022.10.002 |
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