Current diagnostic tests for viral respiratory infections provide limited clinical value and therefore rarely ordered by physicians. A rapid and accurate diagnosis enables early and effective therapeutic intervention for both common infectious pathogens and biothreat agents. The goal of this proposal is to demonstrate the feasibility of a nucleic acid-based multiplex test for the detection and quantification of the most frequently encountered viral pathogens in the respiratory tract, namely influenza A & B, parainfluenza 1,2, & 3, and respiratory syncytial virus. Conserved nucleotide sequences within each viral genome will be identified as targets for RT-PCR amplification using real-time TaqMan(R) chemistry. Each set of primers is optimized for a multiplex reaction format and will amplify with equal efficiency and similar dynamic range. The amplified products are then quantified on the Luminex xMAPTM platform, a bead-based hybridization technology capable of detecting up to 100 targets simultaneously. The unique sequence of the Taqman probe is incorporated as the capture probe on the Luminex beads. This approach combines the exquisite sensitivity of PCR with a rapid, multiplex detection system. The ultimate goal is to provide a cost-effective platform that can simultaneously and efficiently detect and quantify all types of potential pathogens.
