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Application of purified recombinant antigenic spike fragments to the diagnosis of avian infectious bronchitis virus infection

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Abstract

The spike (S) protein, containing two subunits, S1 and S2, is the major immunity-eliciting antigen of avian infectious bronchitis virus (IBV), a highly contagious disease of chickens. Several immunogenic regions, mainly located within the S1 subunit, have been identified. Nonetheless, these immune-dominant regions were defined using selected monoclonal antibodies or using a short peptide approach that involves only certain limited regions of the S protein. In addition, some immune-dominant regions are located in hypervariable regions (HVRs) which are not present in all serotypes. Hence, the aim of this study was to determine a broader range of antigenic regions that have strong antibody eliciting ability; these could then be applied for development of an IBV-diagnostic tool. Initially, the S1 and part of the S2 subunit protein (24–567 amino acids) were expressed as five fragments in prokaryotic system. The antigenicity was confirmed using IBV immunized sera. Performance of the S subfragments was evaluated by ELISA using a panel of field chicken sera with known IBV titres determined by a commercial kit. This indicated that, among the five antigenic recombinant proteins, the region S-E showed the highest specificity and sensitivity, namely 95.38 % and 96.29 %, respectively. The κ value for the in-house ELISA using the S-E fragment compared to a commercial kit was 0.9172, indicating a high agreement between these two methods. As region S-E harbors strong immunogenicity within the spike protein, it has the potential to be exploited as an antigen when developing a cost-effective ELISA-based diagnosis tool.

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Acknowledgments

This study was partly supported by the Bureau of Animal and Plant Health Inspection and Quarantine, the Council of Agriculture, grant number: 96–1.1.5-B1(5).

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Authors and Affiliations

  1. Department of Veterinary Medicine, College of Veterinary Medicine, National Chung Hsing University, 250 Kuo Kuang Road, Taichung, 402, Taiwan

    Kuan-Hsun Lin, Ai-Ping Hsu, Tien-Jye Chang & Jui-Hung Shien

  2. Department of Medical Laboratory Science and Biotechnology, Central Taiwan University of Science and Technology, Taichung, Taiwan

    Chuen-Fu Lin

  3. Graduate Institute of Microbiology and Public Health, College of Veterinary Medicine, National Chung Hsing University, 250 Kuo Kuang Road, Taichung, 402, Taiwan

    Shyan-Song Chiou, Chao-Chin Chang & Wei-Li Hsu

  4. Council of Agriculture, Animal Health Research Institute, 376 Chung-Cheng Road, Tamsui, Taipei, 251, Taiwan

    Ai-Ping Hsu & Min-Shiuh Lee

Authors
  1. Kuan-Hsun Lin
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  2. Chuen-Fu Lin
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  3. Shyan-Song Chiou
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  4. Ai-Ping Hsu
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  5. Min-Shiuh Lee
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  6. Chao-Chin Chang
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  7. Tien-Jye Chang
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  8. Jui-Hung Shien
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  9. Wei-Li Hsu
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Correspondence to Jui-Hung Shien or Wei-Li Hsu.

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Lin, KH., Lin, CF., Chiou, SS. et al. Application of purified recombinant antigenic spike fragments to the diagnosis of avian infectious bronchitis virus infection. Appl Microbiol Biotechnol 95, 233–242 (2012). https://doi.org/10.1007/s00253-012-4143-8

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  • DOI: https://doi.org/10.1007/s00253-012-4143-8

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