Skip to main content

Rotaviruses

  • Protocol
  • First Online:
Diagnostic Virology Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 665))

Abstract

Rotaviruses can be detected easily, and methods have been developed to visualise their characteristic morphology, to detect rotavirus proteins through immunological methods or the virus genome, either directly by polyacrylamide gel electrophoresis or after reverse transcription of the viral RNA and amplification by PCR. The abundance of virus particles found in clinical samples during an acute infection makes the detection of rotavirus proteins, mainly VP6, the method of choice for virus detection. Molecular methods are generally reserved for the characterisation of a diverse population of viruses circulating in many mammalian species. Characterisation methods have been developed to determine diversity within genes encoding viral structural proteins, VP4, VP7, and VP6 and the non-structural protein and viral enterotoxin, NSP4. The combined use of the detection and characterisation methods described in this chapter allows novel rotavirus strains resulting from genetic reassortment among common strains, reassortment among animal and human strains and zoonotic strains to be identified. Also, strains in which diversity is generated through the accumulation of point mutations during virus replication are identified. The development of safe and effective rotavirus vaccines necessitates the detection and characterisation of rotaviruses of genomic and antigenic diversity circulating in both the human and animal populations.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Parashar, U.D., et al. (2006) Rotavirus and severe childhood diarrhea. Emerg Infect Dis 12(2), 304–306.

    Article  PubMed  Google Scholar 

  2. Adah, M.I., et al. (1997) Nigerian rotavirus serotype G8 could not be typed by PCR due to nucleotide mutation at the 3′ end of the primer binding site. Arch Virol 142(9), 1881–1887.

    Article  PubMed  CAS  Google Scholar 

  3. Iturriza-Gómara, M., et al. (2000) Diversity within the VP4 gene of rotavirus P[8] strains: implications for reverse transcription-PCR genotyping. J Clin Microbiol 38(2), 898–901.

    PubMed  Google Scholar 

  4. Iturriza-Gomara, M., Kang, G., and Gray, J.J. (2004) Rotavirus genotyping: keeping up with an evolving population of human rotaviruses. J Med Virol 31(4), 259–265.

    CAS  Google Scholar 

  5. Banerjee, I., et al. (2007) Molecular characterization of G11P[25] and G3P[3] human rotavirus strains associated with asymptomatic infection in South India. J Med Virol 79(11), 1768–1774.

    Article  PubMed  CAS  Google Scholar 

  6. Iturriza Gomara, M., et al. (2004) Characterization of G10P[11] rotaviruses causing acute gastroenteritis in neonates and infants in Vellore, India. J Clin Microbiol 42(6), 2541–2547.

    Article  PubMed  Google Scholar 

  7. Iturriza Gómara, M., et al. (2001) Amino acid substitution within the VP7 protein of G2 rotavirus strains associated with failure to serotype. J Clin Microbiol 39(10), 3796–3798.

    Article  Google Scholar 

  8. Banerjee, I., et al. (2007) Modification of rotavirus multiplex RT-PCR for the detection of G12 strains based on characterization of emerging G12 rotavirus strains from South India. J Med Virol 79(9), 1413–1421.

    Article  PubMed  CAS  Google Scholar 

  9. Iturriza-Gomara, M., et al. (2003) Evidence for genetic linkage between the gene segments encoding NSP4 and VP6 proteins in common and reassortant human rotavirus strains. J Clin Microbiol 41(8), 3566–3573.

    Article  PubMed  CAS  Google Scholar 

  10. Ciarlet, M., et al. (2000) Species specificity and interspecies relatedness of NSP4 genetic groups by comparative NSP4 sequence analyses of animal rotaviruses. Arch Virol 145(2), 371–383.

    Article  PubMed  CAS  Google Scholar 

  11. Iturriza Gomara, M., et al. (2002) Molecular characterization of VP6 genes of human rotavirus isolates: correlation of genogroups with subgroups and evidence of independent segregation. J Virol 76(13), 6596–6601.

    Article  PubMed  Google Scholar 

  12. Amar, C.F., et al. (2007) Detection by PCR of eight groups of enteric pathogens in 4,627 faecal samples: re-examination of the English case-control Infectious Intestinal Disease Study (1993–1996). Eur J Clin Microbiol Infect Dis 26(5), 311–323.

    Article  PubMed  CAS  Google Scholar 

  13. Iturriza-Gómara, M., et al. (2000) Molecular epidemiology of human group A rotavirus infections in the UK between 1995 and 1998. J Clin Microbiol 38, 4394–4401.

    PubMed  Google Scholar 

  14. Gouvea, V., et al. (1990) Polymerase chain reaction amplification and typing of rotavirus nucleic acid from stool specimens. J Clin Microbiol 28(2), 276–282.

    PubMed  CAS  Google Scholar 

  15. Simmonds, M.K., et al. (2008) New oligonucleotide primers for P-typing of rotavirus strains: strategies for typing previously untypeable strains. J Clin Virol 42, 368–373.

    Article  PubMed  CAS  Google Scholar 

  16. Gentsch, J.R., et al. (1992) Identification of group A rotavirus gene 4 types by polymerase chain reaction. J Clin Microbiol 30(6), 1365–1373.

    PubMed  CAS  Google Scholar 

  17. Boom, R., et al. (1990) Rapid and simple method for the purification of nucleic acids. J Clin Microbiol 28, 495–503.

    PubMed  CAS  Google Scholar 

  18. Matthijnssens, J., et al. (2008) Recom­mendations for the classification of group A rotaviruses using all 11genomic RNA segments. Arch Virol 153(8), 1621–1629.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2010 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Gray, J., Iturriza-Gómara, M. (2010). Rotaviruses. In: Stephenson, J., Warnes, A. (eds) Diagnostic Virology Protocols. Methods in Molecular Biology, vol 665. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-817-1_18

Download citation

  • DOI: https://doi.org/10.1007/978-1-60761-817-1_18

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60761-816-4

  • Online ISBN: 978-1-60761-817-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics