Tonje Castberg, Øivind Enger and Gunnar Bratbak
Department of Microbiology, University of Bergen, Jahnebakken 5,
N-5020 Bergen, Norway
Current methods for detection and quantification of viruses are based either on titration, electron or epifluorescence microscopy. Titration requires an established host-virus system and time-consuming culturing, while microscopy lacks potential for specific detection. Ecological studies of algal viruses require reliable, specific techniques for detection and quantification.. To study the ecology of Phaeocystis pouchetii - PpV host virus system we are working on a competitive PCR approach, applying an internal standard template that allows specific and sensitive detection of the virus both in algal cultures and sea water samples. To develop a specific PCR for the Phaeocystis virus (PpV) we have used degenerate primers specific for phycodnaviridae, and sequenced the obtained PCR product to design specific primers for PpV. This approach also allows us to compare the sequenced strains with other well described algal viruses. This highly versatile methodological approach may be applied also to other culturable vira and possibly also to viral systems not yet isolated.