Development of RT-qPCR assays for the specific identification of two major genotypes of avian infectious bronchitis virus
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...
| Main Authors: | , , , , , , , , , , , |
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| Format: | info:eu-repo/semantics/article |
| Language: | Inglés |
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2017
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| Online Access: | 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 |
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| author | 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_browse | Banda, Alejandro Craig, María Isabel Grecco, Sofía Hernández, Diego Hernández, Martín Marandino, Ana Panzera, Yanina Pérez, Ruben Techera, Claudia Tomás, Gonzalo Vagnozzi, Ariel Eduardo Vera, Federico Sebastian |
| 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_sort | Marandino, Ana |
| collection | INTA Digital |
| 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 |
| format | info:eu-repo/semantics/article |
| id | INTA1224 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2017 |
| publishDateRange | 2017 |
| publishDateSort | 2017 |
| record_format | dspace |
| spelling | INTA12242018-01-22T14:30:15Z Development of RT-qPCR assays for the specific identification of two major genotypes of avian infectious bronchitis virus 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 Enfermedades de los Animales Virus Bronquitis Infecciosa Aviar Genotipos Aves de Corral Animal Diseases Avian Infectious Bronchitis Virus PCR Genotypes Poultry 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 2017-09-14T17:08:05Z 2017-09-14T17:08:05Z 2016 info:eu-repo/semantics/article info:ar-repo/semantics/artículo info:eu-repo/semantics/acceptedVersion 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 eng info:eu-repo/semantics/restrictedAccess application/pdf Journal of virological methods 235 : 21-25. (September 2016) |
| spellingShingle | Enfermedades de los Animales Virus Bronquitis Infecciosa Aviar Genotipos Aves de Corral Animal Diseases Avian Infectious Bronchitis Virus PCR Genotypes Poultry 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 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 |
| 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_short | 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 |
| topic | Enfermedades de los Animales Virus Bronquitis Infecciosa Aviar Genotipos Aves de Corral Animal Diseases Avian Infectious Bronchitis Virus PCR Genotypes Poultry |
| 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 |
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