Development of an RT-qPCR assay for the specific detection of a distinct genetic lineage of the infectious bursal disease virus

Tomás Custodio, Gonzalo Martín - Hernández, Martín - Marandino, Ana - Techera, Claudia - Grecco Patiño, Sofía - Hernández López, Diego - Banda, Alejandro - Panzera Crespo, Yanina - Pérez Crossa, Ruben Gustavo

Resumen:

The infectious bursal disease virus (IBDV) is a major health threat to the world’s poultry industry despite intensive controls including proper biosafety practices and vaccination. IBDV (Avibirnavirus, Birnaviridae) is a non-enveloped virus with a bisegmented double-stranded RNA genome. The virus is traditionally classified into classic, variant and very virulent strains, each with different epidemiological relevance and clinical implications. Recently, a novel worldwide spread genetic lineage was described and denoted as distinct (d) IBDV. Here, we report the development and validation of a reverse transcription-quantitative polymerase chain reaction (RT-qPCR) assay for the specific detection of dIBDVs in the global poultry industry. The assay employs a TaqMan-MGB probe that hybridizes with a unique molecular signature of dIBDV. The assay successfully detected all the assessed strains belonging to the dIBDV genetic lineage, showing high specificity and absence of cross-reactivity with non-dIBDVs, IBDVnegative samples and other common avian viruses. Using serial dilutions of in vitro-transcribed RNA we obtained acceptable PCR efficiencies and determination coefficients, and relatively small intra- and inter-assay variability. The assay demonstrated a wide dynamic range between 103 and 108 RNA copies/reaction. This rapid, specific and quantitative assay is expected to improve IBDV surveillance and control worldwide and to increase our understanding of the molecular epidemiology of this economically detrimental poultry pathogen.


Detalles Bibliográficos
2017
IBDV
RT-qPCR
TaqMan-MGB probe
distinct IBDV
IBD
Inglés
Universidad de la República
COLIBRI
https://hdl.handle.net/20.500.12008/34394
Acceso abierto
Licencia Creative Commons Atribución (CC - By 4.0)
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author Tomás Custodio, Gonzalo Martín
author2 Hernández, Martín
Marandino, Ana
Techera, Claudia
Grecco Patiño, Sofía
Hernández López, Diego
Banda, Alejandro
Panzera Crespo, Yanina
Pérez Crossa, Ruben Gustavo
author2_role author
author
author
author
author
author
author
author
author_facet Tomás Custodio, Gonzalo Martín
Hernández, Martín
Marandino, Ana
Techera, Claudia
Grecco Patiño, Sofía
Hernández López, Diego
Banda, Alejandro
Panzera Crespo, Yanina
Pérez Crossa, Ruben Gustavo
author_role author
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http://localhost:8080/xmlui/bitstream/20.500.12008/34394/1/10.108003079457.2016.1228827.pdf
collection COLIBRI
dc.contributor.filiacion.none.fl_str_mv Tomás Custodio Gonzalo Martín, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología.
Hernández Martín, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología.
Marandino Ana, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología.
Techera Claudia, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología.
Grecco Patiño Sofía, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología.
Hernández López Diego, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología.
Banda Alejandro, Mississippi State University (Estados Unidos).
Panzera Crespo Yanina, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología.
Pérez Crossa Ruben Gustavo, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología.
dc.creator.none.fl_str_mv Tomás Custodio, Gonzalo Martín
Hernández, Martín
Marandino, Ana
Techera, Claudia
Grecco Patiño, Sofía
Hernández López, Diego
Banda, Alejandro
Panzera Crespo, Yanina
Pérez Crossa, Ruben Gustavo
dc.date.accessioned.none.fl_str_mv 2022-10-26T16:54:27Z
dc.date.available.none.fl_str_mv 2022-10-26T16:54:27Z
dc.date.issued.none.fl_str_mv 2017
dc.description.abstract.none.fl_txt_mv The infectious bursal disease virus (IBDV) is a major health threat to the world’s poultry industry despite intensive controls including proper biosafety practices and vaccination. IBDV (Avibirnavirus, Birnaviridae) is a non-enveloped virus with a bisegmented double-stranded RNA genome. The virus is traditionally classified into classic, variant and very virulent strains, each with different epidemiological relevance and clinical implications. Recently, a novel worldwide spread genetic lineage was described and denoted as distinct (d) IBDV. Here, we report the development and validation of a reverse transcription-quantitative polymerase chain reaction (RT-qPCR) assay for the specific detection of dIBDVs in the global poultry industry. The assay employs a TaqMan-MGB probe that hybridizes with a unique molecular signature of dIBDV. The assay successfully detected all the assessed strains belonging to the dIBDV genetic lineage, showing high specificity and absence of cross-reactivity with non-dIBDVs, IBDVnegative samples and other common avian viruses. Using serial dilutions of in vitro-transcribed RNA we obtained acceptable PCR efficiencies and determination coefficients, and relatively small intra- and inter-assay variability. The assay demonstrated a wide dynamic range between 103 and 108 RNA copies/reaction. This rapid, specific and quantitative assay is expected to improve IBDV surveillance and control worldwide and to increase our understanding of the molecular epidemiology of this economically detrimental poultry pathogen.
dc.format.extent.es.fl_str_mv 8 h.
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dc.identifier.citation.es.fl_str_mv Tomás Custodio, G, Hernández, M, Marandino, A, [y otros autores] "Development of an RT-qPCR assay for the specific detection of a distinct genetic lineage of the infectious bursal disease virus". Avian Pathology. [en línea] 2017, 46(2): 150–156. 8 h.
dc.identifier.doi.none.fl_str_mv 10.1080/03079457.2016.1228827
dc.identifier.issn.none.fl_str_mv 0307-9457
dc.identifier.uri.none.fl_str_mv https://hdl.handle.net/20.500.12008/34394
dc.language.iso.none.fl_str_mv en_US
eng
dc.publisher.es.fl_str_mv Taylor and Francis Ltd.
dc.relation.ispartof.es.fl_str_mv Avian Pathology, 2017, 46(2): 150–156.
dc.rights.license.none.fl_str_mv Licencia Creative Commons Atribución (CC - By 4.0)
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
dc.rights.uri.*.fl_str_mv An error occurred getting the license - uri.
An error occurred getting the license - uri.
dc.source.none.fl_str_mv reponame:COLIBRI
instname:Universidad de la República
instacron:Universidad de la República
dc.subject.es.fl_str_mv IBDV
RT-qPCR
TaqMan-MGB probe
distinct IBDV
IBD
dc.title.none.fl_str_mv Development of an RT-qPCR assay for the specific detection of a distinct genetic lineage of the infectious bursal disease virus
dc.type.es.fl_str_mv Artículo
dc.type.none.fl_str_mv info:eu-repo/semantics/article
dc.type.version.none.fl_str_mv info:eu-repo/semantics/publishedVersion
description The infectious bursal disease virus (IBDV) is a major health threat to the world’s poultry industry despite intensive controls including proper biosafety practices and vaccination. IBDV (Avibirnavirus, Birnaviridae) is a non-enveloped virus with a bisegmented double-stranded RNA genome. The virus is traditionally classified into classic, variant and very virulent strains, each with different epidemiological relevance and clinical implications. Recently, a novel worldwide spread genetic lineage was described and denoted as distinct (d) IBDV. Here, we report the development and validation of a reverse transcription-quantitative polymerase chain reaction (RT-qPCR) assay for the specific detection of dIBDVs in the global poultry industry. The assay employs a TaqMan-MGB probe that hybridizes with a unique molecular signature of dIBDV. The assay successfully detected all the assessed strains belonging to the dIBDV genetic lineage, showing high specificity and absence of cross-reactivity with non-dIBDVs, IBDVnegative samples and other common avian viruses. Using serial dilutions of in vitro-transcribed RNA we obtained acceptable PCR efficiencies and determination coefficients, and relatively small intra- and inter-assay variability. The assay demonstrated a wide dynamic range between 103 and 108 RNA copies/reaction. This rapid, specific and quantitative assay is expected to improve IBDV surveillance and control worldwide and to increase our understanding of the molecular epidemiology of this economically detrimental poultry pathogen.
eu_rights_str_mv openAccess
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identifier_str_mv Tomás Custodio, G, Hernández, M, Marandino, A, [y otros autores] "Development of an RT-qPCR assay for the specific detection of a distinct genetic lineage of the infectious bursal disease virus". Avian Pathology. [en línea] 2017, 46(2): 150–156. 8 h.
0307-9457
10.1080/03079457.2016.1228827
instacron_str Universidad de la República
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reponame_str COLIBRI
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repository.name.fl_str_mv COLIBRI - Universidad de la República
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Licencia Creative Commons Atribución (CC - By 4.0)
spelling Tomás Custodio Gonzalo Martín, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología.Hernández Martín, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología.Marandino Ana, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología.Techera Claudia, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología.Grecco Patiño Sofía, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología.Hernández López Diego, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología.Banda Alejandro, Mississippi State University (Estados Unidos).Panzera Crespo Yanina, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología.Pérez Crossa Ruben Gustavo, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología.2022-10-26T16:54:27Z2022-10-26T16:54:27Z2017Tomás Custodio, G, Hernández, M, Marandino, A, [y otros autores] "Development of an RT-qPCR assay for the specific detection of a distinct genetic lineage of the infectious bursal disease virus". Avian Pathology. [en línea] 2017, 46(2): 150–156. 8 h.0307-9457https://hdl.handle.net/20.500.12008/3439410.1080/03079457.2016.1228827The infectious bursal disease virus (IBDV) is a major health threat to the world’s poultry industry despite intensive controls including proper biosafety practices and vaccination. IBDV (Avibirnavirus, Birnaviridae) is a non-enveloped virus with a bisegmented double-stranded RNA genome. The virus is traditionally classified into classic, variant and very virulent strains, each with different epidemiological relevance and clinical implications. Recently, a novel worldwide spread genetic lineage was described and denoted as distinct (d) IBDV. Here, we report the development and validation of a reverse transcription-quantitative polymerase chain reaction (RT-qPCR) assay for the specific detection of dIBDVs in the global poultry industry. The assay employs a TaqMan-MGB probe that hybridizes with a unique molecular signature of dIBDV. The assay successfully detected all the assessed strains belonging to the dIBDV genetic lineage, showing high specificity and absence of cross-reactivity with non-dIBDVs, IBDVnegative samples and other common avian viruses. Using serial dilutions of in vitro-transcribed RNA we obtained acceptable PCR efficiencies and determination coefficients, and relatively small intra- and inter-assay variability. The assay demonstrated a wide dynamic range between 103 and 108 RNA copies/reaction. This rapid, specific and quantitative assay is expected to improve IBDV surveillance and control worldwide and to increase our understanding of the molecular epidemiology of this economically detrimental poultry pathogen.Submitted by Farías Verónica (vfarias@fcien.edu.uy) on 2022-10-25T14:33:56Z No. of bitstreams: 2 license_rdf: 19875 bytes, checksum: 9fdbed07f52437945402c4e70fa4773e (MD5) 10.108003079457.2016.1228827.pdf: 1145466 bytes, checksum: 3f88e9417d332131075dcdf7750fcfb9 (MD5)Approved for entry into archive by Faget Cecilia (lfaget@fcien.edu.uy) on 2022-10-25T14:46:54Z (GMT) No. of bitstreams: 2 license_rdf: 19875 bytes, checksum: 9fdbed07f52437945402c4e70fa4773e (MD5) 10.108003079457.2016.1228827.pdf: 1145466 bytes, checksum: 3f88e9417d332131075dcdf7750fcfb9 (MD5)Made available in DSpace by Luna Fabiana (fabiana.luna@seciu.edu.uy) on 2022-10-26T16:54:27Z (GMT). No. of bitstreams: 2 license_rdf: 19875 bytes, checksum: 9fdbed07f52437945402c4e70fa4773e (MD5) 10.108003079457.2016.1228827.pdf: 1145466 bytes, checksum: 3f88e9417d332131075dcdf7750fcfb9 (MD5) Previous issue date: 20178 h.application/pdfen_USengTaylor and Francis Ltd.Avian Pathology, 2017, 46(2): 150–156.Las obras depositadas en el Repositorio se rigen por la Ordenanza de los Derechos de la Propiedad Intelectual de la Universidad de la República.(Res. Nº 91 de C.D.C. de 8/III/1994 – D.O. 7/IV/1994) y por la Ordenanza del Repositorio Abierto de la Universidad de la República (Res. Nº 16 de C.D.C. de 07/10/2014)An error occurred getting the license - uri.An error occurred getting the license - uri.info:eu-repo/semantics/openAccessLicencia Creative Commons Atribución (CC - By 4.0)IBDVRT-qPCRTaqMan-MGB probedistinct IBDVIBDDevelopment of an RT-qPCR assay for the specific detection of a distinct genetic lineage of the infectious bursal disease virusArtículoinfo:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionreponame:COLIBRIinstname:Universidad de la Repúblicainstacron:Universidad de la RepúblicaTomás Custodio, Gonzalo MartínHernández, MartínMarandino, AnaTechera, ClaudiaGrecco Patiño, SofíaHernández López, DiegoBanda, AlejandroPanzera Crespo, YaninaPérez Crossa, Ruben GustavoLICENSElicense.txtlicense.txttext/plain; charset=utf-84267http://localhost:8080/xmlui/bitstream/20.500.12008/34394/5/license.txt6429389a7df7277b72b7924fdc7d47a9MD55CC-LICENSElicense_urllicense_urltext/plain; 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- Universidad de la Repúblicafalse
spellingShingle Development of an RT-qPCR assay for the specific detection of a distinct genetic lineage of the infectious bursal disease virus
Tomás Custodio, Gonzalo Martín
IBDV
RT-qPCR
TaqMan-MGB probe
distinct IBDV
IBD
status_str publishedVersion
title Development of an RT-qPCR assay for the specific detection of a distinct genetic lineage of the infectious bursal disease virus
title_full Development of an RT-qPCR assay for the specific detection of a distinct genetic lineage of the infectious bursal disease virus
title_fullStr Development of an RT-qPCR assay for the specific detection of a distinct genetic lineage of the infectious bursal disease virus
title_full_unstemmed Development of an RT-qPCR assay for the specific detection of a distinct genetic lineage of the infectious bursal disease virus
title_short Development of an RT-qPCR assay for the specific detection of a distinct genetic lineage of the infectious bursal disease virus
title_sort Development of an RT-qPCR assay for the specific detection of a distinct genetic lineage of the infectious bursal disease virus
topic IBDV
RT-qPCR
TaqMan-MGB probe
distinct IBDV
IBD
url https://hdl.handle.net/20.500.12008/34394