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

描述(申请人提供)：过去出现过流感大流行毒株，将来无疑也会出现。许多呼吸道病毒，包括常见的流感病毒株和大流行性流感病毒株，都具有基本上相同的全身症状。公共卫生和临床关注的一个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的格式，可以同时分析包括流感在内的一组呼吸道病毒，但也可以表征流感的H和N基因，以区分毒株(例如H5N1)。HT-FilmArray采用一次性塑料，在其中执行一系列无缝的生化操作，包括：核酸制备、反转录、使用两阶段嵌套多路复用过程的聚合酶链式反应，以及通过高分辨率熔体剖析确认产品身份。套式聚合酶链式反应虽然非常敏感，但由于容易受到扩增子污染，其应用受到了限制。HT-FilmArray包含嵌套程序，从而实现了其最大的优势(灵敏度)，同时消除了其最大的弱点(扩增子污染)。使用高分辨率熔体剖面图来确认积极的结果。熔体分析快速确定曲线形状和熔化温度作为第二层分析，以加强阳性结果并消除假阳性结果。在大流行的情况下，实验室之间的沟通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，以便采取措施遏制疫情。建议的是一个软件模块，允许HT-FilmArray用户通过安全的互联网连接进行通信和共享数据，即使在地理位置偏远的位置也是如此。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，并允许共享和分析独立的数据集。数据挖掘模块将允许一起分析结果，并为数据比较提供灵活的格式。数据共享/数据挖掘将使公共卫生官员能够使用来自多台远程HT-FilmArray仪器的数据来监测病毒活动，并根据这些数据做出合理的决策，以控制疫情。虽然这项建议的重点是流感，但在每年的呼吸道病毒流行期间，HT-FilmArray将作为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.