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

Authors
  • Marandino, Ana1
  • Tomás, Gonzalo1
  • Hernández, Martín1
  • Panzera, Yanina1
  • Craig, María Isabel2
  • Vagnozzi, Ariel2
  • Vera, Federico3
  • Techera, Claudia1
  • Grecco, Sofía1
  • Banda, Alejandro4
  • Hernández, Diego1
  • Pérez, Ruben5
  • 1 Sección Genética Evolutiva, Instituto de Biología, Facultad de Ciencias, Universidad de la República, Iguá 4225, 11400 Montevideo, Uruguay. , (Uruguay)
  • 2 Instituto de Virología, CICVyA, INTA-Castelar, CC 25 (1712) Castelar, Buenos Aires, Argentina. , (Argentina)
  • 3 Laboratorio Sanidad Aviar, INTA- E.E.A, Concepción del Uruguay, Entre Ríos, Argentina. , (Argentina)
  • 4 Poultry Research and Diagnostic Laboratory, College of Veterinary Medicine, Mississippi State University, Pearl, MS 39288, USA.
  • 5 Sección Genética Evolutiva, Instituto de Biología, Facultad de Ciencias, Universidad de la República, Iguá 4225, 11400 Montevideo, Uruguay. Electronic address: [email protected] , (Uruguay)
Type
Published Article
Journal
Journal of virological methods
Publication Date
Sep 01, 2016
Volume
235
Pages
21–25
Identifiers
DOI: 10.1016/j.jviromet.2016.05.007
PMID: 27181213
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

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. Copyright © 2016 Elsevier B.V. All rights reserved.

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