Influenza virus causes a pandemic disease of the respiratory tract that results in significant human morbidity and mortality. Diagnosis of influenza virus infection during the winter months makes up a large component of the clinical virology workload. The goal of this application is to develop transgenic cell lines that facilitate the detection of influenza virus (IV) in clinical specimens. Previously, we developed and patented methods for detecting and quantitating other RNA viruses such as Sindbis virus, a positive-strand RNA virus and respiratory syncytial virus (RSV), a negative-strand RNA virus. The RSV system, while very useful as a research tool, is too complicated in its present format to be useful in a diagnostic test for clinical virology laboratories. Recently, we began working to develop an influenza virus detection system. We constructed a plasmid in which a polymerase I promoter drives transcription of an influenza A virus artificial genomic RNA segment that carries a firefly luciferase gene. Interestingly, we have found that infection of cells transfected with this eDNA exhibited a high level of luciferase whereas uninfected cells expressed background levels. We plan to build on this result and make equivalent influenza B artificial genomic RNA. We will then use these constructs to generate stable cell lines that express a reporter gene only in response to influenza A and/or influenza B virus infection. Such cell lines will form the basis of an influenza detection test which will be developed in Phase II in collaboration with Diagnostic Hybrids, Inc. Athens, OH, and modeled after an existing FDA approved test for herpes simplex virus (ELVIS(r)-HSV).
Thesaurus Terms: Orthomyxoviridae, cell line, diagnosis design /evaluation, diagnostic test, reporter gene, transfection clinical trial, gene expression, influenza, influenzavirus A, influenzavirus B, luciferin monooxygenase biotechnology, clinical research, human subject, patient oriented research, tissue /cell culture
