HT-Film-Array: a system to assess respiratory viruses with emphasis on influenza

描述（由申请人提供）：流感大流行毒株在过去已经出现，毫无疑问在未来也会出现。许多呼吸道病毒，包括常见的流感病毒株和大流行病毒株，表现出基本相同的全身症状。一个公共卫生和临床问题是区分感染大流行性流感或普通呼吸道病毒的患者，这样做需要一个敏感的诊断系统。HT-FilmArray技术提供了一种密集格式，可同时检测一组呼吸道病毒（包括流感病毒），但也可表征流感病毒H和N基因，以区分毒株（例如H5 N1）。HT-FilmArray采用一次性塑料，可进行无缝的生化操作，包括：核酸制备、逆转录、使用2阶段嵌套多重过程的PCR以及通过高分辨率熔解曲线确认产品同一性。巢式PCR虽然灵敏度高，但由于容易受到扩增子污染，因此使用有限。HT-FilmArray包含嵌套程序，从而实现其最大优势（灵敏度），同时消除其最大弱点（扩增子污染）。使用高分辨率熔融分析来确认阳性结果。熔融分析可快速确定曲线形状和熔融温度，作为第二层分析，以加强阳性结果并消除假阳性结果。在大流行情况下，实验室之间的通信将使临床医生能够更好地诊断患者，并为公共卫生官员提供信息，以采取措施遏制疫情。提出的软件模块允许HT- FilmArray用户（即使在地理位置偏远的地方）通过安全的互联网连接进行通信和共享数据。该数据共享模块将使特权用户能够从远程位置实时查看运行情况，并共享和分析独立的数据集。数据挖掘模块将允许对结果进行综合分析，并提供灵活的数据比较格式。数据共享/数据挖掘将使公共卫生官员能够使用来自多个远程HT-FilmArray仪器的数据监测病毒活动，并根据数据做出合理的决策以遏制疫情。虽然该提案的重点是流感，但HT-FilmArray作为每年呼吸道病毒流行期间评估患者的标准工具同样有用。

DESCRIPTION (provided by applicant): Pandemic strains of influenza have arisen in the past and will unquestionably arise in the future. Numerous respiratory viruses, including common influenza strains and pandemic strains, present with essentially identical generalized symptoms. A public health and clinical concern is distinguishing between patients infected with pandemic influenza or a common respiratory virus, to do so requires a sensitive diagnostic system. HT-FilmArray technology provides a dense format to concurrently assay for a panel of respiratory viruses including influenza but to also characterize the influenza H and N genes to differentiate strains (e.g. H5N1). HT-FilmArray employs a plastic disposable in which is performed a seamless progression of biochemical manipulations including: nucleic acid preparation, reverse transcription, PCR using a 2-stage nested multiplex process, and confirmation of product identity by high-resolution melt profiling. Nested PCR while exquisitely sensitive, has seen limited use as it is prone to amplicon contamination. HT-FilmArray contains the nested procedure such that its greatest strength (sensitivity) is realized while its greatest weakness (amplicon contamination) is eliminated. High-resolution melt profiling is used to confirm positive results. Melt profiling rapidly determines curve shape and melting temperature as a 2nd tier analysis to re-enforce positive results and to eliminate false positive results. In a pandemic scenario, communication between laboratories will enable clinicians to better diagnose patients and provide public health officials information to take steps that will contain the outbreak. Proposed is a software module allowing HT- FilmArray users, even in geographically remote locations, to communicate and share data over a secure internet connection. This data sharing module will enable privileged users to view runs in real-time from remote locations and for independent data sets to be shared and analyzed together. A data mining module will allow results to be analyzed together and provide a flexible format for data comparison. Data sharing/data mining will allow public health officials to monitor viral activity using data from multiple distantly located HT-FilmArray instruments and based upon the data make sound decisions to contain an outbreak. While this proposal is focused upon influenza, HT-FilmArray will be equally useful as a standard tool to assess patients during yearly respiratory virus epidemics.