Development of RT-qPCR assays for the specific identification of two major genotypes of avian infectious bronchitis virus

Autores
Marandino, Ana; Tomás, Gonzalo; Hernández, Martín; Panzera, Yanina; Craig, Marí­a Isabel; Vagnozzi, Ariel Eduardo; Vera, Federico Sebastian; Techera, Claudia; Grecco, Sofía; Banda, Alejandro; Hernández, Diego; Pérez, Ruben
Año de publicación
2016
Idioma
inglés
Tipo de recurso
artículo
Estado
versión aceptada
Descripción
Infectious bronchitis virus (Gammacoronavirus, Coronaviridae) is a genetically variable RNA virus (27.6kb) that causes one of the most persistent respiratory disease in poultry. The virus is classified in genotypes with different epidemiological relevance and clinical implications. The present study reports the development and validation of specific RT-qPCR assays for the detection of two major IBV genotypes: South America I (SAI) and Asia/South America II (A/SAII). The SAI genotype is an exclusive and widespread South American lineage while the A/SAII genotype is distributed in Asia, Europe and South America. Both identification assays employ TaqMan probes that hybridize with unique sequences in the spike glycoprotein gene. The assays successfully detected all the assessed strains belonging to both genotypes, showing high specificity and absence of cross-reactivity. Using serial dilutions of in vitro-transcribed RNA we obtained acceptable determination coefficients, PCR efficiencies and relatively small intra- and inter-assay variability. The assays demonstrated a wide dynamic range between 10(1)-10(7) and 10(2)-10(7) RNA copies/reaction for SAI and A/SAII strains, respectively. The possibility to characterize a large number of samples in a rapid, sensitive and reproducible way makes these techniques suitable tools for routine testing, IBV control, and epidemiological research in poultry
Inst.de Virología
Fil: Marandino, Ana. Universidad de la República. Facultad de Ciencias. Instituto de Biología. Sección Genética Evolutiva; Uruguay
Fil: Tomás, Gonzalo. Universidad de la República. Facultad de Ciencias. Instituto de Biología. Sección Genética Evolutiva; Uruguay
Fil: Hernández, Martín. Universidad de la República. Facultad de Ciencias. Instituto de Biología. Sección Genética Evolutiva; Uruguay
Fil: Panzera, Yanina. Universidad de la República. Facultad de Ciencias. Instituto de Biología. Sección Genética Evolutiva; Uruguay
Fil: Craig, Marí­a Isabel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina
Fil: Vagnozzi, Ariel Eduardo. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina
Fil: Vera, Federico Sebastian. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Concepción del Uruguay. Laboratorio Sanidad Aviar; Argentina
Fil: Techera, Claudia. Universidad de la República. Facultad de Ciencias. Instituto de Biología. Sección Genética Evolutiva; Uruguay
Fil: Grecco, Sofía. Universidad de la República. Facultad de Ciencias. Instituto de Biología. Sección Genética Evolutiva; Uruguay
Fil: Banda, Alejandro. Mississippi State University. College of Veterinary Medicine. Poultry Research and Diagnostic Laboratory; Estados Unidos
Fil: Hernández, Diego. Universidad de la República. Facultad de Ciencias. Instituto de Biología. Sección Genética Evolutiva; Uruguay
Fil: Pérez, Ruben. Universidad de la República. Facultad de Ciencias. Instituto de Biología. Sección Genética Evolutiva; Uruguay
Fuente
Journal of virological methods 235 : 21-25. (September 2016)
Materia
Enfermedades de los Animales
Virus Bronquitis Infecciosa Aviar
Genotipos
Aves de Corral
Animal Diseases
Avian Infectious Bronchitis Virus
PCR
Genotypes
Poultry
Nivel de accesibilidad
acceso restringido
Condiciones de uso
Repositorio
INTA Digital (INTA)
Institución
Instituto Nacional de Tecnología Agropecuaria
OAI Identificador
oai:localhost:20.500.12123/1224
Acceder
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oai_identifier_str oai:localhost:20.500.12123/1224
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network_name_str INTA Digital (INTA)
spelling Development of RT-qPCR assays for the specific identification of two major genotypes of avian infectious bronchitis virusMarandino, AnaTomás, GonzaloHernández, MartínPanzera, YaninaCraig, Marí­a IsabelVagnozzi, Ariel EduardoVera, Federico SebastianTechera, ClaudiaGrecco, SofíaBanda, AlejandroHernández, DiegoPérez, RubenEnfermedades de los AnimalesVirus Bronquitis Infecciosa AviarGenotiposAves de CorralAnimal DiseasesAvian Infectious Bronchitis VirusPCRGenotypesPoultryInfectious bronchitis virus (Gammacoronavirus, Coronaviridae) is a genetically variable RNA virus (27.6kb) that causes one of the most persistent respiratory disease in poultry. The virus is classified in genotypes with different epidemiological relevance and clinical implications. The present study reports the development and validation of specific RT-qPCR assays for the detection of two major IBV genotypes: South America I (SAI) and Asia/South America II (A/SAII). The SAI genotype is an exclusive and widespread South American lineage while the A/SAII genotype is distributed in Asia, Europe and South America. Both identification assays employ TaqMan probes that hybridize with unique sequences in the spike glycoprotein gene. The assays successfully detected all the assessed strains belonging to both genotypes, showing high specificity and absence of cross-reactivity. Using serial dilutions of in vitro-transcribed RNA we obtained acceptable determination coefficients, PCR efficiencies and relatively small intra- and inter-assay variability. The assays demonstrated a wide dynamic range between 10(1)-10(7) and 10(2)-10(7) RNA copies/reaction for SAI and A/SAII strains, respectively. The possibility to characterize a large number of samples in a rapid, sensitive and reproducible way makes these techniques suitable tools for routine testing, IBV control, and epidemiological research in poultryInst.de VirologíaFil: Marandino, Ana. Universidad de la República. Facultad de Ciencias. Instituto de Biología. Sección Genética Evolutiva; UruguayFil: Tomás, Gonzalo. Universidad de la República. Facultad de Ciencias. Instituto de Biología. Sección Genética Evolutiva; UruguayFil: Hernández, Martín. Universidad de la República. Facultad de Ciencias. Instituto de Biología. Sección Genética Evolutiva; UruguayFil: Panzera, Yanina. Universidad de la República. Facultad de Ciencias. Instituto de Biología. Sección Genética Evolutiva; UruguayFil: Craig, Marí­a Isabel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; ArgentinaFil: Vagnozzi, Ariel Eduardo. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; ArgentinaFil: Vera, Federico Sebastian. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Concepción del Uruguay. Laboratorio Sanidad Aviar; ArgentinaFil: Techera, Claudia. Universidad de la República. Facultad de Ciencias. Instituto de Biología. Sección Genética Evolutiva; UruguayFil: Grecco, Sofía. Universidad de la República. Facultad de Ciencias. Instituto de Biología. Sección Genética Evolutiva; UruguayFil: Banda, Alejandro. Mississippi State University. College of Veterinary Medicine. Poultry Research and Diagnostic Laboratory; Estados UnidosFil: Hernández, Diego. Universidad de la República. Facultad de Ciencias. Instituto de Biología. Sección Genética Evolutiva; UruguayFil: Pérez, Ruben. Universidad de la República. Facultad de Ciencias. Instituto de Biología. Sección Genética Evolutiva; Uruguay2017-09-14T17:08:05Z2017-09-14T17:08:05Z2016info:eu-repo/semantics/articleinfo:eu-repo/semantics/acceptedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/1224http://www.sciencedirect.com/science/article/pii/S01660934163003250166-0934 (Print)1879-0984 (Online)https://doi.org/10.1016/j.jviromet.2016.05.007Journal of virological methods 235 : 21-25. (September 2016)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo:eu-repo/semantics/restrictedAccess2025-10-16T09:29:00Zoai:localhost:20.500.12123/1224instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2025-10-16 09:29:01.17INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
dc.title.none.fl_str_mv Development of RT-qPCR assays for the specific identification of two major genotypes of avian infectious bronchitis virus
title Development of RT-qPCR assays for the specific identification of two major genotypes of avian infectious bronchitis virus
spellingShingle Development of RT-qPCR assays for the specific identification of two major genotypes of avian infectious bronchitis virus
Marandino, Ana
Enfermedades de los Animales
Virus Bronquitis Infecciosa Aviar
Genotipos
Aves de Corral
Animal Diseases
Avian Infectious Bronchitis Virus
PCR
Genotypes
Poultry
title_short Development of RT-qPCR assays for the specific identification of two major genotypes of avian infectious bronchitis virus
title_full Development of RT-qPCR assays for the specific identification of two major genotypes of avian infectious bronchitis virus
title_fullStr Development of RT-qPCR assays for the specific identification of two major genotypes of avian infectious bronchitis virus
title_full_unstemmed Development of RT-qPCR assays for the specific identification of two major genotypes of avian infectious bronchitis virus
title_sort Development of RT-qPCR assays for the specific identification of two major genotypes of avian infectious bronchitis virus
dc.creator.none.fl_str_mv Marandino, Ana
Tomás, Gonzalo
Hernández, Martín
Panzera, Yanina
Craig, Marí­a Isabel
Vagnozzi, Ariel Eduardo
Vera, Federico Sebastian
Techera, Claudia
Grecco, Sofía
Banda, Alejandro
Hernández, Diego
Pérez, Ruben
author Marandino, Ana
author_facet Marandino, Ana
Tomás, Gonzalo
Hernández, Martín
Panzera, Yanina
Craig, Marí­a Isabel
Vagnozzi, Ariel Eduardo
Vera, Federico Sebastian
Techera, Claudia
Grecco, Sofía
Banda, Alejandro
Hernández, Diego
Pérez, Ruben
author_role author
author2 Tomás, Gonzalo
Hernández, Martín
Panzera, Yanina
Craig, Marí­a Isabel
Vagnozzi, Ariel Eduardo
Vera, Federico Sebastian
Techera, Claudia
Grecco, Sofía
Banda, Alejandro
Hernández, Diego
Pérez, Ruben
author2_role author
author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Enfermedades de los Animales
Virus Bronquitis Infecciosa Aviar
Genotipos
Aves de Corral
Animal Diseases
Avian Infectious Bronchitis Virus
PCR
Genotypes
Poultry
topic Enfermedades de los Animales
Virus Bronquitis Infecciosa Aviar
Genotipos
Aves de Corral
Animal Diseases
Avian Infectious Bronchitis Virus
PCR
Genotypes
Poultry
dc.description.none.fl_txt_mv Infectious bronchitis virus (Gammacoronavirus, Coronaviridae) is a genetically variable RNA virus (27.6kb) that causes one of the most persistent respiratory disease in poultry. The virus is classified in genotypes with different epidemiological relevance and clinical implications. The present study reports the development and validation of specific RT-qPCR assays for the detection of two major IBV genotypes: South America I (SAI) and Asia/South America II (A/SAII). The SAI genotype is an exclusive and widespread South American lineage while the A/SAII genotype is distributed in Asia, Europe and South America. Both identification assays employ TaqMan probes that hybridize with unique sequences in the spike glycoprotein gene. The assays successfully detected all the assessed strains belonging to both genotypes, showing high specificity and absence of cross-reactivity. Using serial dilutions of in vitro-transcribed RNA we obtained acceptable determination coefficients, PCR efficiencies and relatively small intra- and inter-assay variability. The assays demonstrated a wide dynamic range between 10(1)-10(7) and 10(2)-10(7) RNA copies/reaction for SAI and A/SAII strains, respectively. The possibility to characterize a large number of samples in a rapid, sensitive and reproducible way makes these techniques suitable tools for routine testing, IBV control, and epidemiological research in poultry
Inst.de Virología
Fil: Marandino, Ana. Universidad de la República. Facultad de Ciencias. Instituto de Biología. Sección Genética Evolutiva; Uruguay
Fil: Tomás, Gonzalo. Universidad de la República. Facultad de Ciencias. Instituto de Biología. Sección Genética Evolutiva; Uruguay
Fil: Hernández, Martín. Universidad de la República. Facultad de Ciencias. Instituto de Biología. Sección Genética Evolutiva; Uruguay
Fil: Panzera, Yanina. Universidad de la República. Facultad de Ciencias. Instituto de Biología. Sección Genética Evolutiva; Uruguay
Fil: Craig, Marí­a Isabel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina
Fil: Vagnozzi, Ariel Eduardo. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina
Fil: Vera, Federico Sebastian. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Concepción del Uruguay. Laboratorio Sanidad Aviar; Argentina
Fil: Techera, Claudia. Universidad de la República. Facultad de Ciencias. Instituto de Biología. Sección Genética Evolutiva; Uruguay
Fil: Grecco, Sofía. Universidad de la República. Facultad de Ciencias. Instituto de Biología. Sección Genética Evolutiva; Uruguay
Fil: Banda, Alejandro. Mississippi State University. College of Veterinary Medicine. Poultry Research and Diagnostic Laboratory; Estados Unidos
Fil: Hernández, Diego. Universidad de la República. Facultad de Ciencias. Instituto de Biología. Sección Genética Evolutiva; Uruguay
Fil: Pérez, Ruben. Universidad de la República. Facultad de Ciencias. Instituto de Biología. Sección Genética Evolutiva; Uruguay
description Infectious bronchitis virus (Gammacoronavirus, Coronaviridae) is a genetically variable RNA virus (27.6kb) that causes one of the most persistent respiratory disease in poultry. The virus is classified in genotypes with different epidemiological relevance and clinical implications. The present study reports the development and validation of specific RT-qPCR assays for the detection of two major IBV genotypes: South America I (SAI) and Asia/South America II (A/SAII). The SAI genotype is an exclusive and widespread South American lineage while the A/SAII genotype is distributed in Asia, Europe and South America. Both identification assays employ TaqMan probes that hybridize with unique sequences in the spike glycoprotein gene. The assays successfully detected all the assessed strains belonging to both genotypes, showing high specificity and absence of cross-reactivity. Using serial dilutions of in vitro-transcribed RNA we obtained acceptable determination coefficients, PCR efficiencies and relatively small intra- and inter-assay variability. The assays demonstrated a wide dynamic range between 10(1)-10(7) and 10(2)-10(7) RNA copies/reaction for SAI and A/SAII strains, respectively. The possibility to characterize a large number of samples in a rapid, sensitive and reproducible way makes these techniques suitable tools for routine testing, IBV control, and epidemiological research in poultry
publishDate 2016
dc.date.none.fl_str_mv 2016
2017-09-14T17:08:05Z
2017-09-14T17:08:05Z
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/acceptedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str acceptedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/20.500.12123/1224
http://www.sciencedirect.com/science/article/pii/S0166093416300325
0166-0934 (Print)
1879-0984 (Online)
https://doi.org/10.1016/j.jviromet.2016.05.007
url http://hdl.handle.net/20.500.12123/1224
http://www.sciencedirect.com/science/article/pii/S0166093416300325
https://doi.org/10.1016/j.jviromet.2016.05.007
identifier_str_mv 0166-0934 (Print)
1879-0984 (Online)
dc.language.none.fl_str_mv eng
language eng
dc.rights.none.fl_str_mv info:eu-repo/semantics/restrictedAccess
eu_rights_str_mv restrictedAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv Journal of virological methods 235 : 21-25. (September 2016)
reponame:INTA Digital (INTA)
instname:Instituto Nacional de Tecnología Agropecuaria
reponame_str INTA Digital (INTA)
collection INTA Digital (INTA)
instname_str Instituto Nacional de Tecnología Agropecuaria
repository.name.fl_str_mv INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuaria
repository.mail.fl_str_mv tripaldi.nicolas@inta.gob.ar
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