Development of an RT-qPCR assay for the specific detection of a distinct genetic lineage of the infectious bursal disease virus
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.
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) |
_version_ | 1807522794474307584 |
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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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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. |
dc.format.mimetype.es.fl_str_mv | application/pdf |
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 |
format | article |
id | COLIBRI_0f22f499b083572fc13f5157b3c73795 |
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 |
institution | Universidad de la República |
instname_str | Universidad de la República |
language | eng |
language_invalid_str_mv | en_US |
network_acronym_str | COLIBRI |
network_name_str | COLIBRI |
oai_identifier_str | oai:colibri.udelar.edu.uy:20.500.12008/34394 |
publishDate | 2017 |
reponame_str | COLIBRI |
repository.mail.fl_str_mv | mabel.seroubian@seciu.edu.uy |
repository.name.fl_str_mv | COLIBRI - Universidad de la República |
repository_id_str | 4771 |
rights_invalid_str_mv | An error occurred getting the license - uri. 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 |