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Quantitative RT-PCR assays for identification and ty** of the Equine encephalosis virus

  • Veterinary Microbiology - Research Paper
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Abstract

Equine encephalosis (EE) is an acute, arthropod-borne, noncontagious, febrile disease of equids. The clinical signs of EE are similar to milder forms of African horse sickness (AHS) and the two diseases can be easily confused. The Equine encephalosis virus (EEV) is a distinct virus species within the genus Orbivirus, family Reoviridae, with ten linear segments of dsRNA genome. Seven distinct serotypes of EEV have been recognised on the basis of sequence analyses of Seg-2. The need for differential diagnosis of similar forms of EE and AHS warranted the development of molecular diagnostic methods for specific detection and identification of EEV. We report the development of quantitative real-time RT-PCR assay for detection of any member of the EEV species targeting the highly conserved EEV Seg-9. Similar serotype-specific qRT-PCR assays were designed for each of the seven EEV serotypes targeting genome Seg-2, encoding the serotype determining VP2 protein. These assays were evaluated using different EEV serotypes and other closely related orbiviruses. They were shown to be EEV virus species–specific, or EEV type–specific capable of detecting 1 to 13 copies of viral RNA in clinical samples. The assays failed to detect RNA from closely related orbiviruses, including AHSV and Peruvian horse sickness virus (PHSV) isolates.

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Acknowledgments

This work was supported by the Biotechnology and Biological Sciences Research Council (BBSRC) (grant no. BB/L004690/1) and Department for Environment, Food and Rural Affairs (Defra) (SE2617) to PM. Department of Biotechnology, Government of India (DBT, GOI) (grant no. BT/IN/Indo-UK/FADH/46/SM/2013.

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Author notes

  1. Manjunatha N. Belaganahalli

    Present address: One Health Institute, School of Veterinary medicine, University of California, Davis, California, 95616, USA

Authors and Affiliations

  1. College of Veterinary Sciences, LLR University of Veterinary and Animal Sciences, Hisar, Haryana, 125 004, India

    Sushila Maan & Narender Singh Maan

  2. The Pirbright Institute, Ash Road, Pirbright, GU24 0NF, UK

    Sushila Maan, Manjunatha N. Belaganahalli, Narender Singh Maan & Peter P. C. Mertens

  3. Department of Animal Biotechnology, College of Veterinary Sciences, Lala Lajpat Rai University of Veterinary and Animal Sciences (LUVAS), Hisar, Haryana, 125 004, India

    Sushila Maan

  4. Deltamune (Pty) Ltd, 248 Jean Avenue, Lyttelton, Centurion, 0140, South Africa

    Abraham C. Potgieter

  5. Chair of Virology, School of Veterinary Medicine and Science, University of Nottingham, Sutton Bonington Campus, LE12 5RD, UK

    Peter P. C. Mertens

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  1. Sushila Maan
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  2. Manjunatha N. Belaganahalli
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  3. Narender Singh Maan
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Correspondence to Sushila Maan or Manjunatha N. Belaganahalli.

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Sushila Maan and Manjunatha N. Belaganahalli are joint first authors

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Maan, S., Belaganahalli, M.N., Maan, N.S. et al. Quantitative RT-PCR assays for identification and ty** of the Equine encephalosis virus. Braz J Microbiol 50, 287–296 (2019). https://doi.org/10.1007/s42770-018-0034-1

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