Su, Ching-Yao, Wang, Shi-Yun, Shie, Jiun-Jie, Jeng, King-Song, Temperton, Nigel J. ORCID: 0000-0002-7978-3815, Fang, Jim-Min, Wong, Chi-Huey and Cheng, Yih-Shyun E. (2008) In vitro evaluation of neuraminidase inhibitors using the neuraminidase-dependent release assay of hemagglutinin-pseudotyped viruses. Antiviral Research, 79 (3). pp. 199-205. ISSN 0166-3542 (doi:https://doi.org/10.1016/j.antiviral.2008.03.002)

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Abstract

For the treatment of influenza virus infections, neuraminidase inhibitors (NAIs) that prevent the release of virus particles have been effective against most influenza strains. Several neuraminidase (NA) assays are available for the evaluation of NAIs. To understand the NAI functions under physiological conditions, assays mimicking viral particle release should be useful. We have constructed retrovirus-based reporter viruses that are pseudotyped with hemagglutinin (HA) glycoprotein by transfection of producer cells using plasmids expressing retroviral gag-pol, influenza HA, NA, and firefly luciferase genes. Similarly to the life cycle of influenza viruses, the release of pseudotype viruses also requires neuraminidase functions. This requirement was used to develop an assay to evaluate NAI activities by measuring inhibition of pseudotype virus production at different NAI concentrations. The pseudotype virus release assay was used to determine the IC50 values of Oseltamivir carboxylate, Zanamivir, and the novel phosphonate congeners of Oseltamivir againstN1 group neuraminidases and their H274Y Oseltamivir carboxylate-resistant mutants. The deduced IC50 values obtained using the release assay correlated with those determined using the fluorogenic substrate 2'-(4-methylumbelliferyl)-α-d-N-acetylneuraminic acid (MUNANA) and also correlated with the infectivity results.

Item Type:	Article
Uncontrolled Keywords:	neuraminidase inhibitor, pseudotype virus, antiviral, viral release assay, drug resistance,
Subjects:	Q Science > QR Microbiology > QR355 Virology R Medicine > RC Internal medicine
Faculty / School / Research Centre / Research Group:	Faculty of Engineering & Science > Medway School of Pharmacy
Related URLs:	Publisher Organisation
Last Modified:	16 Jul 2012 10:02
URI:	http://gala.gre.ac.uk/id/eprint/8560

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