Transgenic indicator cells for influenza virus

流感病毒转基因指示细胞

• 批准号: 7008801
• 负责人: Paul David Olivo
• 金额: $ 30.89万
• 依托单位: APATH, LLC
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-03-01 至 2007-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7008801
• 关键词: Orthomyxoviridae; biotechnology; cell line; clinical research; clinical trials; diagnosis design /evaluation; diagnostic tests; gene expression; human subject; influenza; influenzavirus A; influenzavirus B; luciferin monooxygenase; patient oriented research; reporter genes; tissue /cell culture; transfection

DESCRIPTION (provided by applicant): 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).

描述（由申请人提供）：流感病毒引起呼吸道的大流行疾病，导致人类的发病率明显。在冬季，诊断流感病毒感染构成了临床病毒学工作量的很大一部分。该应用的目的是开发转基因细胞系，以促进临床标本中流感病毒（IV）的检测。以前，我们开发了专利的方法，用于检测和定量其他RNA病毒，例如sindbis病毒，阳性链RNA病毒和呼吸综合病毒（RSV），一种阴性链RNA病毒。 RSV系统虽然作为一种研究工具非常有用，但目前的格式过于复杂，无法在临床病毒学实验室的诊断测试中有用。最近，我们开始努力开发一种流感病毒检测系统。我们构建了一个质粒，其中聚合酶I启动子驱动含有萤火虫荧光素酶基因的流感病毒人工基因组RNA节段的转录。有趣的是，我们发现用这种EDNA转染的细胞感染表现出高水平的荧光素酶，而未感染的细胞表达了背景水平。我们计划以此结果为基础，并产生同等的流感人工基因组RNA。然后，我们将使用这些构建体来产生稳定的细胞系，该细胞系仅对流感和/或流感B病毒感染来表达报告基因。此类细胞系将构成流感检测测试的基础，该测试将与诊断Hybrids，Inc。雅典，俄亥俄州雅典合作在II期开发，并在现有的FDA批准的疱疹病毒（Elvis（R）-HSV）的FDA批准测试后进行建模。