Fluorescence-based Multiplexed Universal Virus Detection Platform

The aim of this study is to develop a multiplexed virus detection platform capable of detecting within three hours the presence of any known virus, genetically modified virus or unidentified virus, in clinical samples like serum. A PCR based technique will be employed to identify the presence of unique viral sequences. In its ultimate form, the proposed system will encompass a simple and rapid sample preparation step followed by PCR, fluorophore tagging (using a highly stable and novel amplifying fluorescent polymer), and hybridization to a virus-specific oligonucleotide microarray printed on the cap of a PCR tube. The system will also allow for the cloning and subsequent sequencing of nucleic acids to establish the presence of unknown viruses. This diagnostic platform will find applications in screening banked blood for the presence of viruses, detecting the presence of unnatural (mutated) viruses used as bio-warfare agents, and detecting the presence of latent viruses in the human system. In Phase 1 we will perform the following objectives: 1) Standardize and optimize the PCR, 2) Test and optimize the sample preparation protocol, 3) Fabricate microarrays, 4) Optimize microarray hybridization, 5) Clone and sequence PCR products, and 6) Evaluate the sensitivity and specificity of the assay.