Inter-laboratory comparison of eleven quantitative or digital PCR assays for detection of proviral bovine leukemia virus in blood samples

Bovine leukemia virus (BLV) is the etiological agent of enzootic bovine leukosis and causes a persistent infection that can leave cattle with no symptoms. Many countries have been able to successfully eradicate BLV through improved detection and management methods. However, with the increasing novel...

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Main Authors: Pluta, Aneta, Jaworski, Juan Pablo, Droscha, Casey, VanderWeele, Sophie, Taxis, Tasia M., Valas, Stephen, Brnić, Dragan, Jungić, Andreja, Ruano, María José, Sánchez, Azucena, Murakami, Kenji, Nakamura, Kurumi, Puentes, Rodrigo, De Brun, María L., Ruiz, Vanesa, Ladera Gómez, Marla Eliana, Lendez, Pamela Anahí, Dolcini, Guillermina Laura, Fernandes Camargos, Marcelo, Fonseca, Antonio, Barua, Subarna, Wang, Chengming, Giza, Aleksandra, Kuźmak, Jacek
Format: Artículo
Language:Inglés
Published: BioMed Central 2025
Subjects:
Online Access:http://hdl.handle.net/20.500.12123/21479
https://bmcvetres.biomedcentral.com/articles/10.1186/s12917-024-04228-z
https://doi.org/10.1186/s12917-024-04228-z
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author Pluta, Aneta
Jaworski, Juan Pablo
Droscha, Casey
VanderWeele, Sophie
Taxis, Tasia M.
Valas, Stephen
Brnić, Dragan
Jungić, Andreja
Ruano, María José
Sánchez, Azucena
Murakami, Kenji
Nakamura, Kurumi
Puentes, Rodrigo
De Brun, María L.
Ruiz, Vanesa
Ladera Gómez, Marla Eliana
Lendez, Pamela Anahí
Dolcini, Guillermina Laura
Fernandes Camargos, Marcelo
Fonseca, Antonio
Barua, Subarna
Wang, Chengming
Giza, Aleksandra
Kuźmak, Jacek
author_browse Barua, Subarna
Brnić, Dragan
De Brun, María L.
Dolcini, Guillermina Laura
Droscha, Casey
Fernandes Camargos, Marcelo
Fonseca, Antonio
Giza, Aleksandra
Jaworski, Juan Pablo
Jungić, Andreja
Kuźmak, Jacek
Ladera Gómez, Marla Eliana
Lendez, Pamela Anahí
Murakami, Kenji
Nakamura, Kurumi
Pluta, Aneta
Puentes, Rodrigo
Ruano, María José
Ruiz, Vanesa
Sánchez, Azucena
Taxis, Tasia M.
Valas, Stephen
VanderWeele, Sophie
Wang, Chengming
author_facet Pluta, Aneta
Jaworski, Juan Pablo
Droscha, Casey
VanderWeele, Sophie
Taxis, Tasia M.
Valas, Stephen
Brnić, Dragan
Jungić, Andreja
Ruano, María José
Sánchez, Azucena
Murakami, Kenji
Nakamura, Kurumi
Puentes, Rodrigo
De Brun, María L.
Ruiz, Vanesa
Ladera Gómez, Marla Eliana
Lendez, Pamela Anahí
Dolcini, Guillermina Laura
Fernandes Camargos, Marcelo
Fonseca, Antonio
Barua, Subarna
Wang, Chengming
Giza, Aleksandra
Kuźmak, Jacek
author_sort Pluta, Aneta
collection INTA Digital
description Bovine leukemia virus (BLV) is the etiological agent of enzootic bovine leukosis and causes a persistent infection that can leave cattle with no symptoms. Many countries have been able to successfully eradicate BLV through improved detection and management methods. However, with the increasing novel molecular detection methods there have been few efforts to standardize these results at global scale. This study aimed to determine the interlaboratory accuracy and agreement of 11 molecular tests in detecting BLV. Each qPCR/ddPCR method varied by target gene, primer design, DNA input and chemistries. DNA samples were extracted from blood of BLV-seropositive cattle and lyophilized to grant a better preservation during shipping to all participants around the globe. Twenty nine out of 44 samples were correctly identified by the 11 labs and all methods exhibited a diagnostic sensitivity between 74 and 100%. Agreement amongst different assays was linked to BLV copy numbers present in samples and the characteristics of each assay (i.e., BLV target sequence). Finally, the mean correlation value for all assays was within the range of strong correlation. This study highlights the importance of continuous need for standardization and harmonization amongst assays and the different participants. The results underscore the need of an international calibrator to estimate the efficiency (standard curve) of the different assays and improve quantitation accuracy. Additionally, this will inform future participants about the variability associated with emerging chemistries, methods, and technologies used to study BLV. Altogether, by improving tests performance worldwide it will positively aid in the eradication efforts.
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institution Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina)
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spelling INTA214792025-02-27T10:05:53Z Inter-laboratory comparison of eleven quantitative or digital PCR assays for detection of proviral bovine leukemia virus in blood samples Pluta, Aneta Jaworski, Juan Pablo Droscha, Casey VanderWeele, Sophie Taxis, Tasia M. Valas, Stephen Brnić, Dragan Jungić, Andreja Ruano, María José Sánchez, Azucena Murakami, Kenji Nakamura, Kurumi Puentes, Rodrigo De Brun, María L. Ruiz, Vanesa Ladera Gómez, Marla Eliana Lendez, Pamela Anahí Dolcini, Guillermina Laura Fernandes Camargos, Marcelo Fonseca, Antonio Barua, Subarna Wang, Chengming Giza, Aleksandra Kuźmak, Jacek Bovine Leukemia Virus Quantitative Polymerase Chain Reaction PCR Virus Leucemia Bovina Reacción en Cadena de Polimerasa Cuantitativa Bovine leukemia virus (BLV) is the etiological agent of enzootic bovine leukosis and causes a persistent infection that can leave cattle with no symptoms. Many countries have been able to successfully eradicate BLV through improved detection and management methods. However, with the increasing novel molecular detection methods there have been few efforts to standardize these results at global scale. This study aimed to determine the interlaboratory accuracy and agreement of 11 molecular tests in detecting BLV. Each qPCR/ddPCR method varied by target gene, primer design, DNA input and chemistries. DNA samples were extracted from blood of BLV-seropositive cattle and lyophilized to grant a better preservation during shipping to all participants around the globe. Twenty nine out of 44 samples were correctly identified by the 11 labs and all methods exhibited a diagnostic sensitivity between 74 and 100%. Agreement amongst different assays was linked to BLV copy numbers present in samples and the characteristics of each assay (i.e., BLV target sequence). Finally, the mean correlation value for all assays was within the range of strong correlation. This study highlights the importance of continuous need for standardization and harmonization amongst assays and the different participants. The results underscore the need of an international calibrator to estimate the efficiency (standard curve) of the different assays and improve quantitation accuracy. Additionally, this will inform future participants about the variability associated with emerging chemistries, methods, and technologies used to study BLV. Altogether, by improving tests performance worldwide it will positively aid in the eradication efforts. Instituto de Virología Fil: Pluta, Aneta. National Veterinary Research Institute. Department of Biochemistry; Polonia Fil: Pluta, Aneta. National Veterinary Research Institute. Department of Omics Analyses; Polonia Fil: Jaworski, Juan Pablo. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas (IVIT); Argentina Fil: Jaworski, Juan Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Droscha, Casey. CentralStar Cooperative; Estados Unidos Fil: VanderWeele, Sophie. CentralStar Cooperative; Estados Unidos Fil: Taxis, Tasia M. Michigan State University. College of Agriculture and Natural Resources. Department of Animal Science; Estados Unidos Fil: Valas, Stephen. French Agency for Food, Environmental and Occupational Health and Safety. Unit Pathology and Welfare of Ruminants. Niort Laboratory; Francia Fil: Brnić, Dragan. Croatian Veterinary Institute; Croacia Fil: Jungić, Andreja. Croatian Veterinary Institute; Croacia Fil: Ruano, María José. Ministry of Agriculture, Fisheries and Food. Laboratorio Central de Veterinaria; España Fil: Sánchez, Azucena. Ministry of Agriculture, Fisheries and Food. Laboratorio Central de Veterinaria; España Fil: Murakami, Kenji. Iwate University. Faculty of Agriculture. Department of Veterinary Sciences; Japón Fil: Nakamura, Kurumi. Iwate University. Faculty of Agriculture. Department of Veterinary Sciences; Japón Fil: Puentes, Rodrigo. Universidad de la República. Unidad de Microbiología. Facultad de Veterinaria. Departamento de Patobiología; Uruguay Fil: De Brun, María L. Universidad de la República. Unidad de Microbiología. Facultad de Veterinaria. Departamento de Patobiología; Uruguay Fil: Ruiz, Vanesa. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas (IVIT); Argentina Fil: Ruiz, Vanesa. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Ladera Gómez, Marla Eliana. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil (CIVETAN). Departamento SAMP. Laboratorio de Virología; Argentina Fil: Ladera Gómez, Marla Eliana. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Ladera Gómez, Marla Eliana. Comisión de Investigaciones Científicas de la provincia de Buenos Aires (CICPBA); Argentina Fil: Lendez, Pamela Anahí. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil (CIVETAN). Departamento SAMP. Laboratorio de Virología; Argentina Fil: Lendez, Pamela Anahí. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Lendez, Pamela Anahí. Comisión de Investigaciones Científicas de la provincia de Buenos Aires (CICPBA); Argentina Fil: Dolcini, Guillermina Laura. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil (CIVETAN). Departamento SAMP. Laboratorio de Virología; Argentina Fil: Dolcini, Guillermina Laura. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Dolcini, Guillermina Laura. Comisión de Investigaciones Científicas de la provincia de Buenos Aires (CICPBA); Argentina Fil: Fernandes Camargos, Marcelo. Laboratório Federal de Defesa Agropecuária de Minas Gerais; Brasil Fil: Fonseca, Antonio. Laboratório Federal de Defesa Agropecuária de Minas Gerais; Brasil Fil: Barua, Subarna. Auburn University. College of Veterinary Medicine. Department of Pathobiology; Estados Unidos Fil: Wang, Chengming. Auburn University. College of Veterinary Medicine. Department of Pathobiology; Estados Unidos Fil: Giza, Aleksandra. National Veterinary Research Institute. Department of Omics Analyses; Polonia Fil: Kuźmak, Jacek. National Veterinary Research Institute. Department of Biochemistry; Polonia 2025-02-27T10:00:08Z 2025-02-27T10:00:08Z 2024-08 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/21479 https://bmcvetres.biomedcentral.com/articles/10.1186/s12917-024-04228-z 1746-6148 https://doi.org/10.1186/s12917-024-04228-z 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 BioMed Central BMC Veterinary Research 20 : 381 (August 2024)
spellingShingle Bovine Leukemia Virus
Quantitative Polymerase Chain Reaction
PCR
Virus Leucemia Bovina
Reacción en Cadena de Polimerasa Cuantitativa
Pluta, Aneta
Jaworski, Juan Pablo
Droscha, Casey
VanderWeele, Sophie
Taxis, Tasia M.
Valas, Stephen
Brnić, Dragan
Jungić, Andreja
Ruano, María José
Sánchez, Azucena
Murakami, Kenji
Nakamura, Kurumi
Puentes, Rodrigo
De Brun, María L.
Ruiz, Vanesa
Ladera Gómez, Marla Eliana
Lendez, Pamela Anahí
Dolcini, Guillermina Laura
Fernandes Camargos, Marcelo
Fonseca, Antonio
Barua, Subarna
Wang, Chengming
Giza, Aleksandra
Kuźmak, Jacek
Inter-laboratory comparison of eleven quantitative or digital PCR assays for detection of proviral bovine leukemia virus in blood samples
title Inter-laboratory comparison of eleven quantitative or digital PCR assays for detection of proviral bovine leukemia virus in blood samples
title_full Inter-laboratory comparison of eleven quantitative or digital PCR assays for detection of proviral bovine leukemia virus in blood samples
title_fullStr Inter-laboratory comparison of eleven quantitative or digital PCR assays for detection of proviral bovine leukemia virus in blood samples
title_full_unstemmed Inter-laboratory comparison of eleven quantitative or digital PCR assays for detection of proviral bovine leukemia virus in blood samples
title_short Inter-laboratory comparison of eleven quantitative or digital PCR assays for detection of proviral bovine leukemia virus in blood samples
title_sort inter laboratory comparison of eleven quantitative or digital pcr assays for detection of proviral bovine leukemia virus in blood samples
topic Bovine Leukemia Virus
Quantitative Polymerase Chain Reaction
PCR
Virus Leucemia Bovina
Reacción en Cadena de Polimerasa Cuantitativa
url http://hdl.handle.net/20.500.12123/21479
https://bmcvetres.biomedcentral.com/articles/10.1186/s12917-024-04228-z
https://doi.org/10.1186/s12917-024-04228-z
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