SBIR-STTR Award

Influenza viruses are major agents of epidemic and pandemic disease with resultant deaths from pneumonia. Rapid detection of type A influenza permits administration of amantadine to contacts and prevents unnecessary use of antibiotics. Because type A influenza may lead to pneumonia in the elderly, this is of special importance in geriatric facilities. Rapid viral detection systems for influenza viruses (types A and B) will be developed based on production of highly avid monoclonal antibodies to the major internal antigens of these viruses, including nucleoprotein and M-protein (Ml). Monoclonal antibodies will also be produced against NS1 or nonstructural protein found in influenza-virus-infected cells. These antigens show little variation among type A influenza viruses and therefore provide stable reagents that are immune to antigenic shift and drift. The monoclonal antibodies will be used in enzyme-linked immunosorbent assays (ELISAs) and other antibody-based systems. Viro Dynamics' current panel of monoclonal antibodies to M-protein of type A influenza viruses permits direct detection of influenza (HiNi and H3N2) in nasopharyngeal specimens; increased sensitivity will permit detection of influenza in specimens containing lower levels of antigen. The long-term goal is the development of simple, colorimetric ELISA detection systems for influenza. The Phase I goal is the development of monoclonal antibodies to the major internal antigens of type B influenza viruses.

Anticipated Results:
Monoclonal antibodies developed in this program will be applied to the development of ELISA-based diagnostic kits for rapid identification and typing of influenza virus isolates, types A and B. These monoclonal antibodies will also be used in time-resolved fluoroimmunoassay and immunofluorescence analysis for viral detection. Ultimately, ELISA diagnostic kits will permit simple, direct detection of influenza viruses (types A and B) in clinical specimens.National Institute of Allergy And Infectious Diseases

Rapid detection systems will be developed for detecting influenza and parainfluenza viruses in clinical specimens based on development of panels of monoclonal antibodies (mabs) which target the major internal, relatively non-variant proteins. Rapid detection of viruses permits identification of the infecting agent in hours rather than days or weeks required with cultivation. Amantadine can be administered for protection against type A influenza, antivirals are becoming available for treatment of infections with other viruses. Detection of increased influenza activity will provide an early signal for increase of vaccine production by manufacturers. M-proteins (MI) and nucleoproteins (NP) will be purified from influenza viruses, types A and B, and parainfluenza viruses, types 1 and 3. These antigens will serve as immunogens for production of hybridomas as well as immunoadsorbents for ELISA (enzyme-linked immunosorbent assay) screening. We have successfully utilized this approach in producing a panel of mabs to M-protein of type A influenza with application to direct detection of H3N2 and HINI influenza viruses in clinical specimens by time-resolved fluoroimmunoassay (TRFIA). MAbs recognizing different antigenic sites and antigens with the highest affinity constants will be combined to produce the most sensitive virus detection systems possible in TRFIA and ELISA systems.Awardee's statement of the potential commercial applications of the research:The ultimate goal is the development of a simple, rapid assay for detection of both influenza and parainfluenza viruses.National Institute of Allergy and Infectious Diseases (NIAID)