Development of a droplet digital PCR assay for quantification of the proviral load of bovine leukemia virus

Autores
De Brun, María L.; Cosme, Bruno; Petersen, Marcos Iván; Alvarez, Irene; Folgueras-Flatschart, Aurea; Flatschart, Roberto; Panei, Carlos Javier; Puentes, Rodrigo
Año de publicación
2022
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
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
Fuente
Journal of Veterinary Diagnostic Investigation 34 (3) : 439-447 (Mayo 2022)
Materia
Bovine Leukaemia Virus
PCR
Virus Leucemia Bovina
Reacción en Cadena de la Polimerasa
Proviral Load
Carga Viral
Nivel de accesibilidad
acceso restringido
Condiciones de uso
Repositorio
INTA Digital (INTA)
Institución
Instituto Nacional de Tecnología Agropecuaria
OAI Identificador
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spelling Development of a droplet digital PCR assay for quantification of the proviral load of bovine leukemia virusDe Brun, María L.Cosme, BrunoPetersen, Marcos IvánAlvarez, IreneFolgueras-Flatschart, AureaFlatschart, RobertoPanei, Carlos JavierPuentes, RodrigoBovine Leukaemia VirusPCRVirus Leucemia BovinaReacción en Cadena de la PolimerasaProviral LoadCarga ViralDroplet 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 VirologíaFil: De Brun, María L. Universidad de la República. Facultad de Veterinaria. Unidad de Microbiología. Instituto de Patobiología; UruguayFil: Cosme, Bruno. Instituto Nacional de Metrología, Calidad y Tecnología (Inmetro); BrasilFil: Petersen, Marcos Iván. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; ArgentinaFil: Alvarez, Irene. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; ArgentinaFil: Alvarez, Irene. Consejo Nacional de Investigaciones Científicas y Tecnológicas; ArgentinaFil: Folgueras-Flatschart, Aurea. Instituto Nacional de Metrología, Calidad y Tecnología (Inmetro); BrasilFil: Flatschart, Roberto. Instituto Nacional de Metrología, Calidad y Tecnología (Inmetro); BrasilFil: Panei, Carlos Javier. Universidad Nacional de la Plata. Facultad de Ciencias Veterinarias. Laboratorio de Virología; ArgentinaFil: Puentes, Rodrigo. Universidad de la República. Facultad de Veterinaria. Unidad de Microbiología. Instituto de Patobiología; UruguaySage Publications2022-08-04T11:15:19Z2022-08-04T11:15:19Z2022-05info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/12492https://journals.sagepub.com/doi/10.1177/104063872210855811943-4936https://doi.org/10.1177/10406387221085581Journal of Veterinary Diagnostic Investigation 34 (3) : 439-447 (Mayo 2022)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo: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 públicainfo:eu-repo/semantics/restrictedAccess2025-09-04T09:49:28Zoai:localhost:20.500.12123/12492instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2025-09-04 09:49:28.508INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
dc.title.none.fl_str_mv 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
spellingShingle Development of a droplet digital PCR assay for quantification of the proviral load of bovine leukemia virus
De Brun, María L.
Bovine Leukaemia Virus
PCR
Virus Leucemia Bovina
Reacción en Cadena de la Polimerasa
Proviral Load
Carga Viral
title_short 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_sort Development of a droplet digital PCR assay for quantification of the proviral load of bovine leukemia virus
dc.creator.none.fl_str_mv De Brun, María L.
Cosme, Bruno
Petersen, Marcos Iván
Alvarez, Irene
Folgueras-Flatschart, Aurea
Flatschart, Roberto
Panei, Carlos Javier
Puentes, Rodrigo
author De Brun, María L.
author_facet De Brun, María L.
Cosme, Bruno
Petersen, Marcos Iván
Alvarez, Irene
Folgueras-Flatschart, Aurea
Flatschart, Roberto
Panei, Carlos Javier
Puentes, Rodrigo
author_role author
author2 Cosme, Bruno
Petersen, Marcos Iván
Alvarez, Irene
Folgueras-Flatschart, Aurea
Flatschart, Roberto
Panei, Carlos Javier
Puentes, Rodrigo
author2_role author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Bovine Leukaemia Virus
PCR
Virus Leucemia Bovina
Reacción en Cadena de la Polimerasa
Proviral Load
Carga Viral
topic Bovine Leukaemia Virus
PCR
Virus Leucemia Bovina
Reacción en Cadena de la Polimerasa
Proviral Load
Carga Viral
dc.description.none.fl_txt_mv 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
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.
publishDate 2022
dc.date.none.fl_str_mv 2022-08-04T11:15:19Z
2022-08-04T11:15:19Z
2022-05
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/20.500.12123/12492
https://journals.sagepub.com/doi/10.1177/10406387221085581
1943-4936
https://doi.org/10.1177/10406387221085581
url http://hdl.handle.net/20.500.12123/12492
https://journals.sagepub.com/doi/10.1177/10406387221085581
https://doi.org/10.1177/10406387221085581
identifier_str_mv 1943-4936
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 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
dc.rights.none.fl_str_mv info:eu-repo/semantics/restrictedAccess
eu_rights_str_mv restrictedAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Sage Publications
publisher.none.fl_str_mv Sage Publications
dc.source.none.fl_str_mv Journal of Veterinary Diagnostic Investigation 34 (3) : 439-447 (Mayo 2022)
reponame:INTA Digital (INTA)
instname:Instituto Nacional de Tecnología Agropecuaria
reponame_str INTA Digital (INTA)
collection INTA Digital (INTA)
instname_str Instituto Nacional de Tecnología Agropecuaria
repository.name.fl_str_mv INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuaria
repository.mail.fl_str_mv tripaldi.nicolas@inta.gob.ar
_version_ 1842341398656843776
score 12.623145

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