Rapid Detection of Enveloped Viruses.
Annual rept. 1 Nov 84-31 Oct 85,
MOUNT SINAI MEDICAL CENTER OF THE CITY OF NEW YORK NEW YORK
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The development of system for direct detection of type A influenza viruses in clinical specimens, either nasal or throat washings, using monoclonal antibodies specific for M-protein as capture antibody is discussed. Monoclonal antibodies to M-protein from several of our hybridoma cell lines have been found to be capable of detecting virus inartificially seeded clinical specimens down to levels of about 10 ng in 100 microliters. As a part of this study, pediatric nasal wash specimens were monitored for the presence of influenza virus through traditional virus isolation techniques to provide a library of clinical specimens. Ten positive specimens were found out of approximately 80 specimens surveyed. All isolates were of the H3N2 strain. Mice were hyperimmunized to purified M-protein and the spleen cells fused with myloma cells to generate additional hybridoma cell lines secreting antibody to M-protein. Hybridomas produced in an earlier fusion were cloned. A number of parameters in the ELISA test system were explored to optimize conditions to obtain increased sensitivity of virus detection through the use of monoclonal antibodies to M-protein. Immunofluorescence analysis of the infected cells has been used as a means of clinical detection of viruses. Experiments designed to produce monoclonal antibodies to specific epitopes of M-protein are also progressing.
- Medicine and Medical Research