Interlaboratory comparison of six real-time PCR assays for detection of bovine leukemia virus proviral DNA

Autores
Jaworski, Juan Pablo; Pluta, A.; Rola Luszczak, M.; McGowan, S.L.; Finnegan, C.; Heenemann, K.; Carignano, Hugo Adrián; Alvarez, Irene; Murakami, K.; Willems, L.; Vahlenkamp, T.W.; Trono, Karina Gabriela; Choudhury, B.; Kuzmak, J.
Año de publicación
2018
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Quantitative real-time PCR (qPCR) is increasingly being used for the detection of bovine leukemia virus (BLV) proviral DNA. Nevertheless, quality control for the validation and standardization of such tests is currently lacking. Therefore, the present study was initiated by three Office International des Epizooties (OIE) reference laboratories and three collaborating laboratories to measure the interlaboratory variability of six already developed and available BLV qPCR assays. For that purpose, an international panel of 58 DNA samples reflecting the dynamic range of the majority of the assays was distributed to six testing centers. Based on qualitative results, the overall agreement among all six laboratories was moderate. However, significant variability in the measurement of the BLV proviral DNA copy number was observed among different laboratories. Quantitative PCR assays, even when performed by experienced staff, can yield large variability in BLV proviral DNA copy numbers without harmonization. Further standardization of different factors (i.e., utilization of unified protocols and unique calibrators) should increase interlaboratory agreement.
Fil: Jaworski, Juan Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina
Fil: Pluta, A.. National Veterinary Research Institute; Polonia
Fil: Rola Luszczak, M.. National Veterinary Research Institute; Polonia. Animal and Plant Health Agency; Reino Unido
Fil: McGowan, S.L.. Animal and Plant Health Agency; Reino Unido
Fil: Finnegan, C.. Animal and Plant Health Agency; Reino Unido
Fil: Heenemann, K.. University of Leipzig; Alemania
Fil: Carignano, Hugo Adrián. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina
Fil: Alvarez, Irene. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina
Fil: Murakami, K.. Iwate University; Japón
Fil: Willems, L.. Université de Liège; Bélgica
Fil: Vahlenkamp, T.W.. University of Leipzig; Alemania
Fil: Trono, Karina Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina
Fil: Choudhury, B.. Animal and Plant Health Agency; Reino Unido
Fil: Kuzmak, J.. National Veterinary Research Institute; Polonia
Materia
BOVINE LEUKEMIA VIRUS (BLV)
INTERLABORATORY RING TRIAL
PROVIRAL DNA
QUANTITATIVE REAL-TIME PCR (QPCR)
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:11336/99978
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/99978
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Interlaboratory comparison of six real-time PCR assays for detection of bovine leukemia virus proviral DNAJaworski, Juan PabloPluta, A.Rola Luszczak, M.McGowan, S.L.Finnegan, C.Heenemann, K.Carignano, Hugo AdriánAlvarez, IreneMurakami, K.Willems, L.Vahlenkamp, T.W.Trono, Karina GabrielaChoudhury, B.Kuzmak, J.BOVINE LEUKEMIA VIRUS (BLV)INTERLABORATORY RING TRIALPROVIRAL DNAQUANTITATIVE REAL-TIME PCR (QPCR)https://purl.org/becyt/ford/4.3https://purl.org/becyt/ford/4Quantitative real-time PCR (qPCR) is increasingly being used for the detection of bovine leukemia virus (BLV) proviral DNA. Nevertheless, quality control for the validation and standardization of such tests is currently lacking. Therefore, the present study was initiated by three Office International des Epizooties (OIE) reference laboratories and three collaborating laboratories to measure the interlaboratory variability of six already developed and available BLV qPCR assays. For that purpose, an international panel of 58 DNA samples reflecting the dynamic range of the majority of the assays was distributed to six testing centers. Based on qualitative results, the overall agreement among all six laboratories was moderate. However, significant variability in the measurement of the BLV proviral DNA copy number was observed among different laboratories. Quantitative PCR assays, even when performed by experienced staff, can yield large variability in BLV proviral DNA copy numbers without harmonization. Further standardization of different factors (i.e., utilization of unified protocols and unique calibrators) should increase interlaboratory agreement.Fil: Jaworski, Juan Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; ArgentinaFil: Pluta, A.. National Veterinary Research Institute; PoloniaFil: Rola Luszczak, M.. National Veterinary Research Institute; Polonia. Animal and Plant Health Agency; Reino UnidoFil: McGowan, S.L.. Animal and Plant Health Agency; Reino UnidoFil: Finnegan, C.. Animal and Plant Health Agency; Reino UnidoFil: Heenemann, K.. University of Leipzig; AlemaniaFil: Carignano, Hugo Adrián. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; ArgentinaFil: Alvarez, Irene. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; ArgentinaFil: Murakami, K.. Iwate University; JapónFil: Willems, L.. Université de Liège; BélgicaFil: Vahlenkamp, T.W.. University of Leipzig; AlemaniaFil: Trono, Karina Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; ArgentinaFil: Choudhury, B.. Animal and Plant Health Agency; Reino UnidoFil: Kuzmak, J.. National Veterinary Research Institute; PoloniaAmerican Society for Microbiology2018-07info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/99978Jaworski, Juan Pablo; Pluta, A.; Rola Luszczak, M.; McGowan, S.L.; Finnegan, C.; et al.; Interlaboratory comparison of six real-time PCR assays for detection of bovine leukemia virus proviral DNA; American Society for Microbiology; Journal of Clinical Microbiology; 56; 7; 7-2018; 1-110095-1137CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://jcm.asm.org/lookup/doi/10.1128/JCM.00304-18info:eu-repo/semantics/altIdentifier/doi/10.1128/JCM.00304-18info:eu-repo/semantics/altIdentifier/url/https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6018327/info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T14:45:47Zoai:ri.conicet.gov.ar:11336/99978instacron: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 14:45:47.561CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Interlaboratory comparison of six real-time PCR assays for detection of bovine leukemia virus proviral DNA
title Interlaboratory comparison of six real-time PCR assays for detection of bovine leukemia virus proviral DNA
spellingShingle Interlaboratory comparison of six real-time PCR assays for detection of bovine leukemia virus proviral DNA
Jaworski, Juan Pablo
BOVINE LEUKEMIA VIRUS (BLV)
INTERLABORATORY RING TRIAL
PROVIRAL DNA
QUANTITATIVE REAL-TIME PCR (QPCR)
title_short Interlaboratory comparison of six real-time PCR assays for detection of bovine leukemia virus proviral DNA
title_full Interlaboratory comparison of six real-time PCR assays for detection of bovine leukemia virus proviral DNA
title_fullStr Interlaboratory comparison of six real-time PCR assays for detection of bovine leukemia virus proviral DNA
title_full_unstemmed Interlaboratory comparison of six real-time PCR assays for detection of bovine leukemia virus proviral DNA
title_sort Interlaboratory comparison of six real-time PCR assays for detection of bovine leukemia virus proviral DNA
dc.creator.none.fl_str_mv Jaworski, Juan Pablo
Pluta, A.
Rola Luszczak, M.
McGowan, S.L.
Finnegan, C.
Heenemann, K.
Carignano, Hugo Adrián
Alvarez, Irene
Murakami, K.
Willems, L.
Vahlenkamp, T.W.
Trono, Karina Gabriela
Choudhury, B.
Kuzmak, J.
author Jaworski, Juan Pablo
author_facet Jaworski, Juan Pablo
Pluta, A.
Rola Luszczak, M.
McGowan, S.L.
Finnegan, C.
Heenemann, K.
Carignano, Hugo Adrián
Alvarez, Irene
Murakami, K.
Willems, L.
Vahlenkamp, T.W.
Trono, Karina Gabriela
Choudhury, B.
Kuzmak, J.
author_role author
author2 Pluta, A.
Rola Luszczak, M.
McGowan, S.L.
Finnegan, C.
Heenemann, K.
Carignano, Hugo Adrián
Alvarez, Irene
Murakami, K.
Willems, L.
Vahlenkamp, T.W.
Trono, Karina Gabriela
Choudhury, B.
Kuzmak, J.
author2_role author
author
author
author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv BOVINE LEUKEMIA VIRUS (BLV)
INTERLABORATORY RING TRIAL
PROVIRAL DNA
QUANTITATIVE REAL-TIME PCR (QPCR)
topic BOVINE LEUKEMIA VIRUS (BLV)
INTERLABORATORY RING TRIAL
PROVIRAL DNA
QUANTITATIVE REAL-TIME PCR (QPCR)
purl_subject.fl_str_mv https://purl.org/becyt/ford/4.3
https://purl.org/becyt/ford/4
dc.description.none.fl_txt_mv Quantitative real-time PCR (qPCR) is increasingly being used for the detection of bovine leukemia virus (BLV) proviral DNA. Nevertheless, quality control for the validation and standardization of such tests is currently lacking. Therefore, the present study was initiated by three Office International des Epizooties (OIE) reference laboratories and three collaborating laboratories to measure the interlaboratory variability of six already developed and available BLV qPCR assays. For that purpose, an international panel of 58 DNA samples reflecting the dynamic range of the majority of the assays was distributed to six testing centers. Based on qualitative results, the overall agreement among all six laboratories was moderate. However, significant variability in the measurement of the BLV proviral DNA copy number was observed among different laboratories. Quantitative PCR assays, even when performed by experienced staff, can yield large variability in BLV proviral DNA copy numbers without harmonization. Further standardization of different factors (i.e., utilization of unified protocols and unique calibrators) should increase interlaboratory agreement.
Fil: Jaworski, Juan Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina
Fil: Pluta, A.. National Veterinary Research Institute; Polonia
Fil: Rola Luszczak, M.. National Veterinary Research Institute; Polonia. Animal and Plant Health Agency; Reino Unido
Fil: McGowan, S.L.. Animal and Plant Health Agency; Reino Unido
Fil: Finnegan, C.. Animal and Plant Health Agency; Reino Unido
Fil: Heenemann, K.. University of Leipzig; Alemania
Fil: Carignano, Hugo Adrián. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina
Fil: Alvarez, Irene. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina
Fil: Murakami, K.. Iwate University; Japón
Fil: Willems, L.. Université de Liège; Bélgica
Fil: Vahlenkamp, T.W.. University of Leipzig; Alemania
Fil: Trono, Karina Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina
Fil: Choudhury, B.. Animal and Plant Health Agency; Reino Unido
Fil: Kuzmak, J.. National Veterinary Research Institute; Polonia
description Quantitative real-time PCR (qPCR) is increasingly being used for the detection of bovine leukemia virus (BLV) proviral DNA. Nevertheless, quality control for the validation and standardization of such tests is currently lacking. Therefore, the present study was initiated by three Office International des Epizooties (OIE) reference laboratories and three collaborating laboratories to measure the interlaboratory variability of six already developed and available BLV qPCR assays. For that purpose, an international panel of 58 DNA samples reflecting the dynamic range of the majority of the assays was distributed to six testing centers. Based on qualitative results, the overall agreement among all six laboratories was moderate. However, significant variability in the measurement of the BLV proviral DNA copy number was observed among different laboratories. Quantitative PCR assays, even when performed by experienced staff, can yield large variability in BLV proviral DNA copy numbers without harmonization. Further standardization of different factors (i.e., utilization of unified protocols and unique calibrators) should increase interlaboratory agreement.
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/11336/99978
Jaworski, Juan Pablo; Pluta, A.; Rola Luszczak, M.; McGowan, S.L.; Finnegan, C.; et al.; Interlaboratory comparison of six real-time PCR assays for detection of bovine leukemia virus proviral DNA; American Society for Microbiology; Journal of Clinical Microbiology; 56; 7; 7-2018; 1-11
0095-1137
CONICET Digital
CONICET
url http://hdl.handle.net/11336/99978
identifier_str_mv Jaworski, Juan Pablo; Pluta, A.; Rola Luszczak, M.; McGowan, S.L.; Finnegan, C.; et al.; Interlaboratory comparison of six real-time PCR assays for detection of bovine leukemia virus proviral DNA; American Society for Microbiology; Journal of Clinical Microbiology; 56; 7; 7-2018; 1-11
0095-1137
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://jcm.asm.org/lookup/doi/10.1128/JCM.00304-18
info:eu-repo/semantics/altIdentifier/doi/10.1128/JCM.00304-18
info:eu-repo/semantics/altIdentifier/url/https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6018327/
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
dc.publisher.none.fl_str_mv American Society for Microbiology
publisher.none.fl_str_mv American Society for Microbiology
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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