An Assay for Rapid and Sensitive Flu Virus Detection

DESCRIPTION (provided by applicant): The specific aims of this Phase I SBIR proposal are to determine the feasibility of a novel detection technology for rapid and sensitive detection of influenza virus. Specifically, we will determine if the manual assay is rapid (about 15 minutes), sensitive (as least 100 times as good as, or better than, an FDA approved antigen test in terms of TCID50 units/mL), reproducible (with CVlt15%) and specific (without false positive results from potentially interfering bacteri al species at concentrations of upto106 cfu/mL). There are five specific aims in the proposed studies, including: 1. Synthesis, purification and qualification of luciferin - 4,7-di-methyl-O-neuraminic acid substrate - Section D1. Our goal is to obta in at least 1 gram of highly purified substrate (gt98% purity by HPLC). 2. Formulation of the detection mix - Section D2. We will prepare a detection mix that contains all necessary reagents for detection of flu viral neuraminidase. The detection mi x will be lyophilized in a single-use detection tube. 3. Assay optimization - Section D3 4. Evaluation of analytical performance - Section D4. Analytical performance of the assay will be evaluated. We will determine the analytical sensitivity for more than 10 strains of human influenza A and human influenza B. 5. Comparison studies and preliminary evaluation using human samples - Section D5. We will perform comparison studies using FDA-approved antigen-based test and RT-PCR assay. In ad dition, we will test some samples collected from human subjects. PUBLIC HEALTH RELVANCE The proposed Phase I SBIR project will evaluate the feasibility of a novel detection technology for rapid and sensitive detection of influenza virus. The techn ology simplifies the detection process by using a master mix which enables one-step detection, i.e., addition of a sample to the master mix followed by light signal measurement, all performed in the same detection tube. The entire process takes 10-15 minut es. Successful development of the proposed influenza virus test will provide a better alternative for rapid flu diagnosis, thereby significantly improving our medical capability for interpandemic influenza and potential pandemic influenza.