Rapid detection and non-subjective characterisation of infectious bronchitis virus isolates using high-resolution melt curve analysis and a mathematical model
AuthorHewson, K; Noormohammadi, AH; Devlin, JM; Mardani, K; Ignjatovic, J
Source TitleArchives of Virology
University of Melbourne Author/sHEWSON, KYLIE; Hadjinoormohammadi, Amir; Devlin, Joanne; Ignjatovic, Jagoda; Mardani, Karim
AffiliationFaculty of Veterinary Science, Veterinary Science
Document TypeJournal Article
CitationsHewson, K., Noormohammadi, A. H., Devlin, J. M., Mardani, K. & Ignjatovic, J. (2009). Rapid detection and non-subjective characterisation of infectious bronchitis virus isolates using high-resolution melt curve analysis and a mathematical model. ARCHIVES OF VIROLOGY, 154 (4), pp.649-660. https://doi.org/10.1007/s00705-009-0357-1.
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© 2009 Springer Verlag. The publisher’s version is restricted access in accordance with the Springer Verlag policy. The original publication is available at http://www.springer.com/springerwiennewyork/medicine/journal/705
Infectious bronchitis virus (IBV) is a coronavirus that causes upper respiratory, renal and/or reproductive diseases with high morbidity in poultry. Classification of IBV is important for implementation of vaccination strategies to control the disease in commercial poultry. Currently, the lengthy process of sequence analysis of the IBV S1 gene is considered the gold standard for IBV strain identification, with a high nucleotide identity (e.g. > or =95%) indicating related strains. However, this gene has a high propensity to mutate and/or undergo recombination, and alone it may not be reliable for strain identification. A real-time polymerase chain reaction (RT-PCR) combined with high-resolution melt (HRM) curve analysis was developed based on the 3'UTR of IBV for rapid detection and classification of IBV from commercial poultry. HRM curves generated from 230 to 435-bp PCR products of several IBV strains were subjected to further analysis using a mathematical model also developed during this study. It was shown that a combination of HRM curve analysis and the mathematical model could reliably group 189 out of 190 comparisons of pairs of IBV strains in accordance with their 3'UTR and S1 gene identities. The newly developed RT-PCR/HRM curve analysis model could detect and rapidly identify novel and vaccine-related IBV strains, as confirmed by S1 gene and 3'UTR nucleotide sequences. This model is a rapid, reliable, accurate and non-subjective system for detection of IBVs in poultry flocks.
Keywordsinfectious bronchitis virus; IBV; poultry; vaccination strategies
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