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Molecular typing of N gene and 3′ untranslated region of IBV field isolates and vaccine strains using RT-PCR and RFLP

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Abstract

Whole nucleocapsid (N) gene and 3′ untranslated region (UTR) of infectious bronchitis virus (IBV) vaccines and Iranian field isolates were amplified using reverse transcription and polymerase chain reaction (RT-PCR). The amplified fragments were subjected to digestion using two restriction endonuclease enzymes, AluI and MnlI. Five different restriction fragment length polymorphism (RFLP) patterns were generated using both enzymes, classifying IBV strains into five similar groups. Based on RT-PCR and RFLP analysis of the N gene and 3′ UTR, IBV vaccine strains H52, H120 and MA5 all from the same serotype generated identical patterns using both enzymes. Vaccine strain and field isolate of 4/91 also had the same pattern distinct from other IBVs. The IB88 vaccine strain and two other field isolates MNS-7862-1 and Ur1/09 had three different RFLP patterns distinguishable from each other and the other IBVs. In conclusion, RT-PCR and RFLP analysis of the N gene and 3′ UTR could be employed as a useful method for differentiating IBV strains especially in cases where S1 gene amplification is not successful because of its highly variable nature among different IBVs.

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Acknowledgments

The authors sincerely thank the institute for bioscience and biotechnology, Urmia University where this investigation was carried out and also thank the deputy dean for research of Urmia University for funding this project.

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Authors and Affiliations

  1. Department of Microbiology, Faculty of Veterinary Medicine, Urmia University, Urmia, West Azerbaijan, Iran

    Raheleh Majdani & Ahmad Morshedi

  2. Department of Food Hygiene and Quality Control, Faculty of Veterinary Medicine, Urmia University, Urmia, West Azerbaijan, Iran

    Karim Mardani

  3. Department of Clinical Science, Faculty of Veterinary Medicine, Tehran University, Tehran, Iran

    Mehdi Vasfi Marandi

  4. Department of Clinical Science, Faculty of Veterinary Medicine, Urmia University, Urmia, West Azerbaijan, Iran

    Alireza Talebi

  5. Faculty of Pharmacy, Tabriz University of Medical Sciences and Health Services, Tabriz, East Azerbaijan, Iran

    Iraj Yazdani

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  1. Raheleh Majdani
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  2. Karim Mardani
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  3. Ahmad Morshedi
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  4. Mehdi Vasfi Marandi
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  5. Alireza Talebi
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  6. Iraj Yazdani
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Correspondence to Raheleh Majdani.

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Majdani, R., Mardani, K., Morshedi, A. et al. Molecular typing of N gene and 3′ untranslated region of IBV field isolates and vaccine strains using RT-PCR and RFLP. Comp Clin Pathol 21, 283–287 (2012). https://doi.org/10.1007/s00580-010-1093-3

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  • DOI: https://doi.org/10.1007/s00580-010-1093-3

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