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Updated: Mar 17, 2026
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Generation, Amplification, and Titration of Recombinant Respiratory Syncytial Viruses
Published on: April 4, 2019
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RSV Growth and Quantification by Microtitration and qRT-PCR Assays
Hayat Caidi1, Jennifer L Harcourt1, Lia M Haynes2
1Gastroenteritis and Respiratory Viruses Laboratory Branch, Division of Viral Diseases, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention (CDC), 1600 Clifton Road NE, Mailstop G-18, Atlanta, GA, 30333, USA.
Methods in Molecular Biology (Clifton, N.J.)
|July 29, 2016
Summary
A new method effectively removes defective interfering particles from respiratory syncytial virus (RSV), enhancing viral preparation. This stabilized RSV remains viable when stored in 25% sucrose at -152°C.
Area of Science:
- Virology
- Molecular Biology
Background:
- Defective interfering viral particles influence viral infection course and reduce virulence.
- Respiratory syncytial virus (RSV) is a significant human pathogen.
Purpose of the Study:
- To develop a simple method for removing defective interfering particles from RSV.
- To optimize the stabilization and storage of high-titer RSV preparations.
Main Methods:
- Limiting dilution technique for removing defective interfering particles.
- Evaluation of sucrose concentrations (25%) for viral stabilization.
- Storage of RSV at -152°C.
- RSV titration, microtitration, quantitative real-time RT-PCR, and immunostaining for characterization.
Main Results:
- A simple limiting dilution method successfully removed defective interfering particles from RSV.
- High-titer viral preparations were obtained from HEp-2 and Vero cell lines.
- RSV demonstrated stability when prepared and stored in 25% sucrose at -152°C.
Conclusions:
- The described method provides a straightforward approach to obtain pure, high-titer RSV.
- Optimized storage conditions ensure the stability and viability of RSV preparations for research and potential therapeutic applications.
