Methods to Study Viruses

This chapter describes methods for growing, purifying, counting, and characterizing viruses. It also provides general principles of diagnostic virology. As obligate intracellular parasites, viruses require cell in which to replicate. The cells must express appropriate receptors and other proteins required by the virus. Cultured cells are often used to study basic steps in virus replication. Viruses can be purified away from cellular proteins and organelles using centrifugation techniques. Most viruses cannot be seen using standard light microscopes, but are often imaged using electron microscopy. Methods that combine image collection and computationally demanding image processing can provide incredible details about virus architecture. Another common way to visualize viruses is to use fluorescent tags or dyes. Although these techniques do not show detailed virus structures, they can be used to follow the progress of a virus through a cell and can provide a direct window into protein–protein interactions required for virus replication. A more indirect method to detect viruses is to look for virally induced changes to cell morphology. A variety of basic biochemical techniques are useful for analyzing viral proteins and nucleic acids. As the viral genomes are relatively simple, they can be manipulated/mutated to study the function of virtually any viral protein. Powerful genetic techniques can also be used to generate “designer” cells or organisms. There are a variety of methods for quantitating viruses. Infectivity assays measure the ability of a virus to productively infect a cell. Techniques that identify specific viral proteins or genomes provide ways to rapidly identify viruses. Some of these assays can be used at the bedside, or in the field. Powerful and inexpensive DNA sequencing technologies are being used to identify new viruses, many of which could not be found by other methods. The challenge is to understand how or if these viruses impact their hosts.