Abstract
Defective interfering viral particles have been reported as important determinants of the course of viral infection, and they can markedly temper the virulence of the infection. Here, we describe a simple method, based on limiting dilution, for the removal of defective interfering particles from RSV. This method results in a high-titer viral preparation from both HEp-2 and Vero cell lines. We evaluated two concentrations of sucrose to stabilize the virus preparation, and demonstrate that RSV is stable when prepared and stored in 25 % sucrose at −152 °C. In addition, this chapter describes some commonly used methods of RSV titration, detection using microtitration and quantitative real-time RT-PCR, and the use of immunostaining for antigenic characterization.
*Authors contributed equally.The findings and conclusions in this report are those of the author(s) and do not necessarily represent the official position of the Centers for Disease Control and Prevention.
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The authors would like to thank Dr. Kathleen Tatti for her critical review of the manuscript and editorial comments.
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Caidi, H., Harcourt, J.L., Haynes, L.M. (2016). RSV Growth and Quantification by Microtitration and qRT-PCR Assays. In: Tripp, R., Jorquera, P. (eds) Human Respiratory Syncytial Virus. Methods in Molecular Biology, vol 1442. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3687-8_2
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DOI: https://doi.org/10.1007/978-1-4939-3687-8_2
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