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Growing poxviruses and determining virus titer.
Girish J Kotwal1, Melissa-Rose Abrahams
1Division of Medical Virology, University of Cape Town, Cape Town, South Africa.
Methods in Molecular Biology (Clifton, N.J.)
|April 29, 2004
Summary
This study details methods for growing and purifying poxviruses, which are cell-associated viruses. Key steps include mechanical cell lysis, sucrose cushion centrifugation, and sucrose gradient purification for large-scale preparation.
Area of Science:
- Virology
- Cell Biology
Background:
- Poxviruses are significant pathogens requiring robust cultivation and purification methods.
- Understanding poxvirus replication is crucial for vaccine development and antiviral research.
Purpose of the Study:
- To describe established methods for the large-scale growth and purification of poxviruses.
- To provide a detailed protocol for researchers working with poxviruses.
Main Methods:
- Poxviruses are cultivated in cell cultures (adherent or suspension) or embryonated hen's eggs.
- Virus isolation involves mechanical lysis of infected cells.
- Semipurification is achieved via centrifugation through a sucrose cushion.
- Further purification employs sucrose gradient ultracentrifugation.
Main Results:
- The described methods enable the preparation of large quantities of purified poxviruses.
- The protocol ensures efficient isolation and purification, maintaining virus integrity.
Conclusions:
- The chapter provides a comprehensive guide to poxvirus propagation and purification.
- These methods are essential for downstream applications such as structural studies and vaccine production.