Using Aptamer Binding Assays to develop a Rapid, DNA Based Test for the Detection of Ranavirus

Ranavirusis an emerging, highly contagious disease amongst amphibians, reptiles and fish. This double stranded-DNA virus has a unique icosahedral shape with a major capsid protein that forms 50% of its total mass. For developing a molecular diagnostic technique for this virus, we chose aptamers (short single stranded DNA sequences) as the target detection probes. Our goal is to generate an aptamer probe through a process called SELEX for Ranavirus Major Capsid protein and Ranavirus whole virion particles. After performing SELEX on both purified recombinant MCP and Virus, about 40 Aptamer sequences were obtained through cloning and sequencing. Using a Nitrocellulose disc Binding Assay approach , three select biotinylated aptamers were subsequently used to determine binding to immobilized MCP. Steptavidin-HorseRadish Peroxidase and color development were used for detection. Details on this assay setup will be presented.