Development of a droplet digital PCR assay for quantification of the proviral load of bovine leukemia virus
Droplet digital PCR (ddPCR) is a highly sensitive tool developed for the detection and quantification of short-sequence variants—a tool that offers unparalleled precision enabling measurement of smaller-fold changes. We describe here the use of ddPCR for the detection of Bovine leukemia virus (BLV)...
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| Format: | Artículo |
| Language: | Inglés |
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Sage Publications
2022
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| Online Access: | http://hdl.handle.net/20.500.12123/12492 https://journals.sagepub.com/doi/10.1177/10406387221085581 https://doi.org/10.1177/10406387221085581 |
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| author | De Brun, María L. Cosme, Bruno Petersen, Marcos Iván Alvarez, Irene Folgueras-Flatschart, Aurea Flatschart, Roberto Panei, Carlos Javier Puentes, Rodrigo |
| author_browse | Alvarez, Irene Cosme, Bruno De Brun, María L. Flatschart, Roberto Folgueras-Flatschart, Aurea Panei, Carlos Javier Petersen, Marcos Iván Puentes, Rodrigo |
| author_facet | De Brun, María L. Cosme, Bruno Petersen, Marcos Iván Alvarez, Irene Folgueras-Flatschart, Aurea Flatschart, Roberto Panei, Carlos Javier Puentes, Rodrigo |
| author_sort | De Brun, María L. |
| collection | INTA Digital |
| description | Droplet digital PCR (ddPCR) is a highly sensitive tool developed for the detection and quantification of short-sequence variants—a tool that offers unparalleled precision enabling measurement of smaller-fold changes. We describe here the use of ddPCR for the detection of Bovine leukemia virus (BLV) DNA provirus. Serum samples and whole blood from experimentally infected sheep and naturally infected cattle were analyzed through ddPCR to detect the BLV gp51 gene, and then compared with serologic and molecular tests. The ddPCR assay was significantly more accurate and sensitive than AGID, ELISA, nested PCR, and quantitative PCR. The limit of detection of ddPCR was 3.3 copies/µL, detecting positive experimentally infected sheep beginning at 6 d post-infection. The ddPCR methodology offers a promising tool for evaluating the BLV proviral load, particularly for the detection of low viral loads. |
| format | Artículo |
| id | INTA12492 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2022 |
| publishDateRange | 2022 |
| publishDateSort | 2022 |
| publisher | Sage Publications |
| publisherStr | Sage Publications |
| record_format | dspace |
| spelling | INTA124922022-08-04T11:22:47Z Development of a droplet digital PCR assay for quantification of the proviral load of bovine leukemia virus De Brun, María L. Cosme, Bruno Petersen, Marcos Iván Alvarez, Irene Folgueras-Flatschart, Aurea Flatschart, Roberto Panei, Carlos Javier Puentes, Rodrigo Bovine Leukaemia Virus PCR Virus Leucemia Bovina Reacción en Cadena de la Polimerasa Proviral Load Carga Viral Droplet digital PCR (ddPCR) is a highly sensitive tool developed for the detection and quantification of short-sequence variants—a tool that offers unparalleled precision enabling measurement of smaller-fold changes. We describe here the use of ddPCR for the detection of Bovine leukemia virus (BLV) DNA provirus. Serum samples and whole blood from experimentally infected sheep and naturally infected cattle were analyzed through ddPCR to detect the BLV gp51 gene, and then compared with serologic and molecular tests. The ddPCR assay was significantly more accurate and sensitive than AGID, ELISA, nested PCR, and quantitative PCR. The limit of detection of ddPCR was 3.3 copies/µL, detecting positive experimentally infected sheep beginning at 6 d post-infection. The ddPCR methodology offers a promising tool for evaluating the BLV proviral load, particularly for the detection of low viral loads. Instituto de Virología Fil: De Brun, María L. Universidad de la República. Facultad de Veterinaria. Unidad de Microbiología. Instituto de Patobiología; Uruguay Fil: Cosme, Bruno. Instituto Nacional de Metrología, Calidad y Tecnología (Inmetro); Brasil Fil: Petersen, Marcos Iván. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; Argentina Fil: Alvarez, Irene. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; Argentina Fil: Alvarez, Irene. Consejo Nacional de Investigaciones Científicas y Tecnológicas; Argentina Fil: Folgueras-Flatschart, Aurea. Instituto Nacional de Metrología, Calidad y Tecnología (Inmetro); Brasil Fil: Flatschart, Roberto. Instituto Nacional de Metrología, Calidad y Tecnología (Inmetro); Brasil Fil: Panei, Carlos Javier. Universidad Nacional de la Plata. Facultad de Ciencias Veterinarias. Laboratorio de Virología; Argentina Fil: Puentes, Rodrigo. Universidad de la República. Facultad de Veterinaria. Unidad de Microbiología. Instituto de Patobiología; Uruguay 2022-08-04T11:15:19Z 2022-08-04T11:15:19Z 2022-05 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/12492 https://journals.sagepub.com/doi/10.1177/10406387221085581 1943-4936 https://doi.org/10.1177/10406387221085581 eng info:eu-repograntAgreement/INTA/2019-PD-E5-I103-001/2019-PD-E5-I103-001/AR./Desarrollo de tecnologías diagnósticas y estudios epidemiológicos para el control de enfermedades que afectan la producción animal y la salud pública info:eu-repo/semantics/restrictedAccess application/pdf Sage Publications Journal of Veterinary Diagnostic Investigation 34 (3) : 439-447 (Mayo 2022) |
| spellingShingle | Bovine Leukaemia Virus PCR Virus Leucemia Bovina Reacción en Cadena de la Polimerasa Proviral Load Carga Viral De Brun, María L. Cosme, Bruno Petersen, Marcos Iván Alvarez, Irene Folgueras-Flatschart, Aurea Flatschart, Roberto Panei, Carlos Javier Puentes, Rodrigo Development of a droplet digital PCR assay for quantification of the proviral load of bovine leukemia virus |
| title | Development of a droplet digital PCR assay for quantification of the proviral load of bovine leukemia virus |
| title_full | Development of a droplet digital PCR assay for quantification of the proviral load of bovine leukemia virus |
| title_fullStr | Development of a droplet digital PCR assay for quantification of the proviral load of bovine leukemia virus |
| title_full_unstemmed | Development of a droplet digital PCR assay for quantification of the proviral load of bovine leukemia virus |
| title_short | Development of a droplet digital PCR assay for quantification of the proviral load of bovine leukemia virus |
| title_sort | development of a droplet digital pcr assay for quantification of the proviral load of bovine leukemia virus |
| topic | Bovine Leukaemia Virus PCR Virus Leucemia Bovina Reacción en Cadena de la Polimerasa Proviral Load Carga Viral |
| url | http://hdl.handle.net/20.500.12123/12492 https://journals.sagepub.com/doi/10.1177/10406387221085581 https://doi.org/10.1177/10406387221085581 |
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