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.
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References
Flint SJ, Enquist W, Racaniello VR, Skalka AM (2009) Virological metyhods in principles of virology. ASM press, Washington, USA
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
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
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
Jia Y (2012) Real-time PCR. In: Methods in cell biology, vol 112. Academic Press, New York, pp 55–68
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
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
Kemeny DM, Challacombe SJ (eds) (1991) ELISA and other solid phase immunoassays: theoretical and practical aspects. John Wiley & Sons, Chichester
Lequin RM (2005) Enzyme immunoassay (EIA)/enzyme-linked immunosorbent assay (ELISA). Clin Chem 51(12):2415–2418
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.
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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
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DOI: https://doi.org/10.1007/978-1-0716-1358-0_17
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