Quantification of bovine leukemia virus proviral DNA using a low-cost real-time polymerase chain reaction

Autores
Petersen Cruceño, Marcos Iván; Alvarez, Irene; Trono, Karina Gabriela; Jaworski, Juan Pablo
Año de publicación
2018
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The detection of bovine leukemia virus (BLV) proviral DNA is an important tool to address whether an animal is infected with BLV. Compared with serological assays, real-time PCR accounts for greater sensitivity and can serve as a confirmatory test for the clarification of inconclusive or discordant serological test results. However, the high cost related to real-time PCR assays has limited their systematic inclusion in BLV surveillance and eradication programs. The aim of the present study was to validate a low-cost quantitative real-time PCR. Interestingly, by using SYBR Green detection dye, we were able to reduce the cost of a single reaction by a factor of 5 compared with most common assays based on the use of fluorogenic probes (i.e., TaqMan technology). This approach allowed a highly sensitive and specific detection and quantification of BLV proviral DNA from purified peripheral blood leukocytes and a milk matrix. Due to its simplicity and low cost, our in-house BLV SYBR quantitative real-time PCR might be used either as a screening or as a confirmatory test in BLV control programs.
Fil: Petersen Cruceño, Marcos Iván. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Alvarez, Irene. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Trono, Karina Gabriela. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Jaworski, Juan Pablo. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Materia
BOVINE LEUKEMIA VIRUS
EPIDEMIOLOGY
PROVIRAL DNA
REAL-TIME POLYMERASE CHAIN REACTION
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/97540
Acceder
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network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Quantification of bovine leukemia virus proviral DNA using a low-cost real-time polymerase chain reactionPetersen Cruceño, Marcos IvánAlvarez, IreneTrono, Karina GabrielaJaworski, Juan PabloBOVINE LEUKEMIA VIRUSEPIDEMIOLOGYPROVIRAL DNAREAL-TIME POLYMERASE CHAIN REACTIONhttps://purl.org/becyt/ford/4.3https://purl.org/becyt/ford/4The detection of bovine leukemia virus (BLV) proviral DNA is an important tool to address whether an animal is infected with BLV. Compared with serological assays, real-time PCR accounts for greater sensitivity and can serve as a confirmatory test for the clarification of inconclusive or discordant serological test results. However, the high cost related to real-time PCR assays has limited their systematic inclusion in BLV surveillance and eradication programs. The aim of the present study was to validate a low-cost quantitative real-time PCR. Interestingly, by using SYBR Green detection dye, we were able to reduce the cost of a single reaction by a factor of 5 compared with most common assays based on the use of fluorogenic probes (i.e., TaqMan technology). This approach allowed a highly sensitive and specific detection and quantification of BLV proviral DNA from purified peripheral blood leukocytes and a milk matrix. Due to its simplicity and low cost, our in-house BLV SYBR quantitative real-time PCR might be used either as a screening or as a confirmatory test in BLV control programs.Fil: Petersen Cruceño, Marcos Iván. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Alvarez, Irene. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Trono, Karina Gabriela. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Jaworski, Juan Pablo. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaElsevier2018-07info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/97540Petersen Cruceño, Marcos Iván; Alvarez, Irene; Trono, Karina Gabriela; Jaworski, Juan Pablo; Quantification of bovine leukemia virus proviral DNA using a low-cost real-time polymerase chain reaction; Elsevier; Journal of Dairy Science; 101; 7; 7-2018; 6366-63740022-0302CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0022030218303503info:eu-repo/semantics/altIdentifier/doi/10.3168/jds.2017-14253info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T15:00:31Zoai:ri.conicet.gov.ar:11336/97540instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-10-15 15:00:32.162CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Quantification of bovine leukemia virus proviral DNA using a low-cost real-time polymerase chain reaction
title Quantification of bovine leukemia virus proviral DNA using a low-cost real-time polymerase chain reaction
spellingShingle Quantification of bovine leukemia virus proviral DNA using a low-cost real-time polymerase chain reaction
Petersen Cruceño, Marcos Iván
BOVINE LEUKEMIA VIRUS
EPIDEMIOLOGY
PROVIRAL DNA
REAL-TIME POLYMERASE CHAIN REACTION
title_short Quantification of bovine leukemia virus proviral DNA using a low-cost real-time polymerase chain reaction
title_full Quantification of bovine leukemia virus proviral DNA using a low-cost real-time polymerase chain reaction
title_fullStr Quantification of bovine leukemia virus proviral DNA using a low-cost real-time polymerase chain reaction
title_full_unstemmed Quantification of bovine leukemia virus proviral DNA using a low-cost real-time polymerase chain reaction
title_sort Quantification of bovine leukemia virus proviral DNA using a low-cost real-time polymerase chain reaction
dc.creator.none.fl_str_mv Petersen Cruceño, Marcos Iván
Alvarez, Irene
Trono, Karina Gabriela
Jaworski, Juan Pablo
author Petersen Cruceño, Marcos Iván
author_facet Petersen Cruceño, Marcos Iván
Alvarez, Irene
Trono, Karina Gabriela
Jaworski, Juan Pablo
author_role author
author2 Alvarez, Irene
Trono, Karina Gabriela
Jaworski, Juan Pablo
author2_role author
author
author
dc.subject.none.fl_str_mv BOVINE LEUKEMIA VIRUS
EPIDEMIOLOGY
PROVIRAL DNA
REAL-TIME POLYMERASE CHAIN REACTION
topic BOVINE LEUKEMIA VIRUS
EPIDEMIOLOGY
PROVIRAL DNA
REAL-TIME POLYMERASE CHAIN REACTION
purl_subject.fl_str_mv https://purl.org/becyt/ford/4.3
https://purl.org/becyt/ford/4
dc.description.none.fl_txt_mv The detection of bovine leukemia virus (BLV) proviral DNA is an important tool to address whether an animal is infected with BLV. Compared with serological assays, real-time PCR accounts for greater sensitivity and can serve as a confirmatory test for the clarification of inconclusive or discordant serological test results. However, the high cost related to real-time PCR assays has limited their systematic inclusion in BLV surveillance and eradication programs. The aim of the present study was to validate a low-cost quantitative real-time PCR. Interestingly, by using SYBR Green detection dye, we were able to reduce the cost of a single reaction by a factor of 5 compared with most common assays based on the use of fluorogenic probes (i.e., TaqMan technology). This approach allowed a highly sensitive and specific detection and quantification of BLV proviral DNA from purified peripheral blood leukocytes and a milk matrix. Due to its simplicity and low cost, our in-house BLV SYBR quantitative real-time PCR might be used either as a screening or as a confirmatory test in BLV control programs.
Fil: Petersen Cruceño, Marcos Iván. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Alvarez, Irene. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Trono, Karina Gabriela. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Jaworski, Juan Pablo. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
description The detection of bovine leukemia virus (BLV) proviral DNA is an important tool to address whether an animal is infected with BLV. Compared with serological assays, real-time PCR accounts for greater sensitivity and can serve as a confirmatory test for the clarification of inconclusive or discordant serological test results. However, the high cost related to real-time PCR assays has limited their systematic inclusion in BLV surveillance and eradication programs. The aim of the present study was to validate a low-cost quantitative real-time PCR. Interestingly, by using SYBR Green detection dye, we were able to reduce the cost of a single reaction by a factor of 5 compared with most common assays based on the use of fluorogenic probes (i.e., TaqMan technology). This approach allowed a highly sensitive and specific detection and quantification of BLV proviral DNA from purified peripheral blood leukocytes and a milk matrix. Due to its simplicity and low cost, our in-house BLV SYBR quantitative real-time PCR might be used either as a screening or as a confirmatory test in BLV control programs.
publishDate 2018
dc.date.none.fl_str_mv 2018-07
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/11336/97540
Petersen Cruceño, Marcos Iván; Alvarez, Irene; Trono, Karina Gabriela; Jaworski, Juan Pablo; Quantification of bovine leukemia virus proviral DNA using a low-cost real-time polymerase chain reaction; Elsevier; Journal of Dairy Science; 101; 7; 7-2018; 6366-6374
0022-0302
CONICET Digital
CONICET
url http://hdl.handle.net/11336/97540
identifier_str_mv Petersen Cruceño, Marcos Iván; Alvarez, Irene; Trono, Karina Gabriela; Jaworski, Juan Pablo; Quantification of bovine leukemia virus proviral DNA using a low-cost real-time polymerase chain reaction; Elsevier; Journal of Dairy Science; 101; 7; 7-2018; 6366-6374
0022-0302
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0022030218303503
info:eu-repo/semantics/altIdentifier/doi/10.3168/jds.2017-14253
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.22299

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