Preparation and application of fluorescently labeled virus particles
Full Citation: Comeau, A. M., and R. T. Noble. 2010. Preparation and application of fluorescently labeled virus particles, p. 19-29. In S. W. Wilhelm, M. G. Weinbauer, and C. A. Suttle [eds.], Manual of Aquatic Viral Ecology. ASLO. [DOI 10.4319/mave.2010.978-0-9845591-0-7.19]
ABSTRACT: Tracing the fate of individual host cells and viruses is a challenging problem for microbial ecology. The emergence of a new assay, using fluorescently labeled viruses (FLVs), offers the promise of a quick and easy method for monitoring host dynamics and virus decay. Using FLVs as probes (FLVPs) for host cells, an assay was optimized for use with SYBR Green I-stained phages and was shown to be an efficient and reliable method for detection of a strain of Vibrio sp. Various microcosm experiments were conducted that demonstrated the utility of the FLVP assay in resolving ecological interactions at the community level. The assay was also used to show that FLVPs, at high enough multiplicities of infection, can directly inhibit viral infection by "titering out" or "coating" the host's cell surface receptors. FLVs can also be used as tracers for studies of virus production and decay. The approach is mathematically similar to the isotope dilution technique, employed in the past to simultaneously measure the release and uptake of ammonium and amino acids. The method can be used to determine rates of viral degradation, production, and turnover for investigations of microbial food webs in aquatic systems.