Molecular characterization and phylogenetic study base on nucleocapsid gene of avian infectious bronchitis viruses isolated from broiler farms, 2014-2015

Authors

1 Department of Poultry Diseases, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

2 Department of Microbiology and Immunology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

3 Department of Research and Production of Poultry Viral Vaccines, Razi Vaccine and Serum Research Institute, Karaj, Iran

Abstract

BACKGROUND: Avian infectious bronchitis (IB) is an economically important poultry disease. The emergence of new infectious bronchitis virus genotypes has complicated IB control programs. Objectives: This is the first comprehensive molecular analysis of the Nucleocapsid (N) gene of Iranian IBVs. Methods: The nucleocapsid gene of ten IBV isolates (which belonges to four different genotypes) was amplified using specific primers. The phylogenetic trees were constructed based on nucleotide and amino acid sequences of “N” gene. Results: IBV genotyping based on “N” gene showed similar IBV classification which was obtained from spike gene analysis and ten isolates  belonged to Massachusetts, QX, 793/B and Variant-2 genotypes. Different strains had 89.97- 99.75% homology in their amino acid sequences. The highest nucleotide sequence similarity was observed between IBKG-1 and IBKG-8 (793/B type IBVs), while the lowest was seen between IBKG-5 and IBKG-9 (QX- type and Variant-2 type) IBV isolates. This low similarity is of an interest because the N protein is highly conserved among different IBV strains. “N” Protein structural analysis revealed that the isolates has 8to 10 alpha helices and 6 to 8 beta sheets. Conclusions: The present study provided basic information to develop recombinant nucleocapsid proteins that are applicable in rapid diagnostic tests and ELISA and recombinant vaccines.

Keywords

References

 
Abreu, J.T., Resende, J.S., Flatschart, R.B., Folgueras-Flatschart, Á.V., Mendes, A.C.R., Martins, N.R., Silva, C.B., Ferreira, M.C., Resende, M. (2006) Molecular analysis of Brazilian infectious bronchitis field isolates by reverse transcription-polymerase chain reaction, restriction fragment length polymorphism, and partial sequencing of the N gene. Avian Dis.  50: 494-501.
Akrami, H., Hedayati, A., Farshian, M., Haqshenas, G. (2013) Development of a recombinant protein-based dot-blot hybridization assay for the detection of antibody to chicken infectious bronchitis virus. Iranian Journal of Veterinary Research. 14: 350-353.
Bande, F., Arshad, S.S., Hair Bejo, M., Moeini, H., Omar, A.R. (2015) Progress and challenges toward the development of vaccines against avian infectious bronchitis. J Immunol Res. 2015: 1-12.
Boursnell, M., Binns, M., Foulds, I., Brown, T. (1985) Sequences of the nucleocapsid genes from two strains of avian infectious bronchitis virus. J Gen Virol. 66: 573-580.
Breslin, J.J., Smith, L.G., Fuller, F.J., Guy, J.S. (1999) Sequence analysis of the turkey coronavirus nucleocapsid protein gene and 3′ untranslated region identifies the virus as a close relative of infectious bronchitis virus. Virus Res. 65: 187-193.
Callison, S.A., Hilt, D.A., Jackwood, M.W. (2005) In vitro analysis of a hammerhead ribozyme targeted to infectious bronchitis virus nucleocapsid mRNA. Avian Dis. 49: 159-163.
Cook, J.K., Jackwood, M., Jones, R.C. (2012)The long view: 40 years of infectious bronchitis research. Avian Pathol. 41: 239-50.
De Wit, J., Cook, J.K. (2014) Factors influencing the outcome of infectious bronchitis vaccination and challenge experiments. Avian Pathol. 43: 485-497.
Han, Z., Zhao, F., Shao, Y., Liu, X., Kong, X., Song, Y., Liu, S. (2013) Fine level epitope mapping and conservation analysis of two novel linear B-cell epitopes of the avian infectious bronchitis coronavirus nucleocapsid protein. Virus Res. 171: 54-64.
Hosseini, H., Bozorgmehri Fard, M.H., Charkhkar, S., Morshed, R. (2015) Epidemiology of Avian Infectious Bronchitis Virus Genotypes in Iran (2010-2014). Avian Dis. 59: 431-5.
Jackwood, M.W. (2012) Review of infectious bronchitis virus around the world. Avian Dis. 56: 634-41.
Lugovskaya, N., Scherbakov, A., Yakovleva, A., Tsyvanyuk, M., Mudrak, N., Drygin, V., Borisov, A. (2006) Detection of antibodies to avian infectious bronchitis virus by a recombinant nucleocapsid protein-based enzyme-linked immunosorbent assay. Journal of Virolo Methods. 135: 292-296.
Najafi, H., Langeroudi, A., Hashemzadeh, M., Karimi, V., Madadgar, O., Ghafouri, S., Maghsoudlo, H., Farahani, R. (2015)  Molecular characterization of infectious bronchitis viruses isolated from broiler chicken farms in Iran, 2014-2015. Arch Virol. 161: 53-62.
Nosrati, M., Tahmorespoor, M., Nassiry, M. (2013) Genetic analysis of three structural proteins in Iranian infectious bronchitis virus isolate. Iranian J Appl Anim Sci. 3: 617-623.
Seo, S.H., Wang, L., Smith, R., Collisson, E. W. (1997) The carboxyl-terminal 120-residue polypeptide of infectious bronchitis virus nucleocapsid induces cytotoxic T lymphocytes and protects chickens from acute infection. J Virol.  71: 7889-94.
Seyfi Abad Shapouri, M., Mayahi, M., Assasi, K., Charkhkar, S. (2004) A survey of the prevalence of infectious bronchitis virus type 4/91 in Iran. Acta Vet Hung. 52: 163-166.
Sheih, H.K., Shein, J.-H., Chou, H.-Y., Shimizhu, Y., Chen, J.-N., Chang, P.-C. (2004) Complete nucleotide sequences of S1 and N genes of infectious bronchitis virus isolated in Japan and Taiwan. Journal Vet Med Sci.  66: 555-558.
Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M., Kumar, S. (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28: 2731-9.
Vasfi-Marandi, M., Bozorgmehrifard, M. (2001) Isolation and identification of infectious bronchitis viruses in chickens between 1997-2000 in Iran . J Facul Vet Med Uni. 56: 119-124.
Williams, A.K., Li, W., Sneed, L.W., Collisson, E.W. (1992) Comparative analyses of the nucleocapsid genes of several strains of infectious bronchitis virus and other coronaviruses. Virus Res. 25: 213-222.
Mardani, K., Morshedi, A., Talebi, A., Vasfi Marandi, M.  (2011) Molecular analysis of the nucleocapsid gene and 3’untranslated region of two infectious Bronchitis Virus field isolates from Iranian poultry farms. Iranian J Vet Med. 5: 53-58.
Journal of Veterinary Research
Volume 71, Issue 4 - Serial Number 4
December 2016
Pages 463-471

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