Skip to main content

High-Throughput Screening Assays for Dengue Antiviral Drug Development

  • Protocol
  • First Online:
Antimicrobial Therapies

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2296))

  • 1464 Accesses

Abstract

Dengue is an arthropod-borne viral disease that has become endemic and a global threat in over 100 countries. The increase in prevalence would require a long-term measure to control outbreaks. Sanofi Pasteur has licensed the tetravalent dengue vaccine (Dengvaxia) in certain dengue endemic countries. However, the efficacy of the vaccine is limited against certain dengue serotypes and can only be used for individuals from the age from 9 to 45 years old. Over the years, there has been intense research conducted on the development of antivirals against dengue virus (DENV) through either inhibiting the virus replication or targeting the host cell mechanism to block the virus entry. However, no approved antiviral drug against dengue is yet available. In this chapter, we describe the dengue antiviral development workflow including (i) prophylactic, (ii) virucidal, and (iii) postinfection assays that are employed in the antiviral drug screening process against DENV. Further, we demonstrate different methods that can be used to enumerate the reduction in virus foci number including foci-forming unit reduction assay (FFURA), estimation of viral RNA copy number through quantitative real-time PCR, and a high-throughput enzyme linked immunosorbent assay (ELISA)-based quantification of virus particles.

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 109.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 139.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 199.99
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. Flint SJ, Enquist W, Racaniello VR, Skalka AM (2009) Virological metyhods in principles of virology. ASM press, Washington, USA

    Google Scholar 

  2. Ramanathan B, Poh CL, Kirk K, McBride WJH, Aaskov J, Grollo L (2016) Synthetic B-cell epitopes eliciting cross-neutralizing antibodies: strategies for future dengue vaccine. PLoS One 11(5):e0155900

    Article  Google Scholar 

  3. Ho JY, Chern JH, Hsieh CF, Liu ST, Liu CJ, Wang YS, Kuo TW, Hsu SJ, Yeh TK, Shih SR, Hsieh PW (2016) In vitro and in vivo studies of a potent capsid-binding inhibitor of enterovirus 71. J Antimicrob Chemother 71(7):1922–1932

    Article  CAS  Google Scholar 

  4. Alker AP, Mwapasa V, Meshnick SR (2004) Rapid real-time PCR genotyping of mutations associated with sulfadoxine-pyrimethamine resistance in Plasmodium falciparum. Antimicrob Agents Chemother 48:2924–2929

    Article  CAS  Google Scholar 

  5. Jia Y (2012) Real-time PCR. In: Methods in cell biology, vol 112. Academic Press, New York, pp 55–68

    Chapter  Google Scholar 

  6. O’Leary JJ, Sheils O, Martin C, Crowley A (2003) TaqMan® technology and real-time polymerase chain reaction. In: Crocker J, Murray PG (eds) Molecular biology in cellular pathology. Wiley, Chichester, p 251

    Chapter  Google Scholar 

  7. Maddocks S, Jenkins R (2017) Chapter 4–quantitative PCR: things to consider. In: Maddocks S, Jenkins R (eds) Understanding PCR. Academic, Boston, pp 45–52

    Chapter  Google Scholar 

  8. Kemeny DM, Challacombe SJ (eds) (1991) ELISA and other solid phase immunoassays: theoretical and practical aspects. John Wiley & Sons, Chichester

    Google Scholar 

  9. Lequin RM (2005) Enzyme immunoassay (EIA)/enzyme-linked immunosorbent assay (ELISA). Clin Chem 51(12):2415–2418

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by the Fundamental Research Grant Scheme (FRGS/1/2018/SKK11/SYUC/03/1) from Ministry of Education, Malaysia and Sunway University Internal Grant (INT-2018-SST-DBS-04) to B.R.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Babu Ramanathan .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2021 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Jabanathan, S.G., Xuan, L.Z., Ramanathan, B. (2021). High-Throughput Screening Assays for Dengue Antiviral Drug Development. In: Barreiro, C., Barredo, JL. (eds) Antimicrobial Therapies. Methods in Molecular Biology, vol 2296. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1358-0_17

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-1358-0_17

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1357-3

  • Online ISBN: 978-1-0716-1358-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics