FilmArray Biothreat Pathogens: An Automated Multiplex PCR Diagnostic System

FilmArray 生物威胁病原体：自动多重 PCR 诊断系统

• 批准号: 7645313
• 负责人: IRENE M OTA
• 金额: $ 139.24万
• 依托单位: BIOFIRE DEFENSE, LLC.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-20 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7645313
• 关键词: Anthrax disease; Bacteria; Biological Assay; Biological Warfare; Bioterrorism; Blood; Categories; Cerebrospinal Fluid; Cerebrum; Cessation of life; Characteristics; Child; Cities; Clinical; Computer software; Critical Illness; DNA; Data; Department of Defense; Detection; Diagnosis; Diagnostic; District of Columbia; Event; Freeze Drying; Fright; Funding; Health; Hospitalization; Hospitals; Hour; Human; Idaho; Individual; Infection; Institutes; Instruction; Kansas; Laboratories; Letters; Life; Magnetic Resonance Imaging; Manuals; Medical; Medical center; Methods; Modification; National Institute of Allergy and Infectious Disease; Nose; Nucleic Acids; Organism; Pathologist; Pathology; Patients; Performance; Polymerase Chain Reaction; Preparation; Process; Public Health; RNA; Reaction; Reagent; Recovery; Reporting; Research Institute; Research Personnel; Risk; Running; Sampling; Screening procedure; Sensitivity and Specificity; Simulate; Sodium Chloride; Solutions; Specificity; Specimen; Staging; Strategic Planning; Surveys; Swab; Symptoms; Syndrome; System; Technology; Temperature; Terrorism; Testing; Time; United States; United States National Institutes of Health; Universities; Upper arm; Viral; Virus; Whole Blood; Writing; base; biodefense; biothreat; cost; density; design; hospital laboratories; instrument; magnetic beads; meetings; microorganism; novel; pathogen; point of care; programs; respiratory; response; seal; software development

DESCRIPTION (provided by applicant): Deliberate acts of terrorism against the United States demonstrated the importance of equipping our nation with the means to defend against these actions. The anthrax letters in 2001 had severe impacts on human health, resulting in 11 hospitalizations, 5 deaths, and fear based on inability to rapidly diagnose. In response to these attacks, Idaho Technology Inc (ITI) created diagnostic systems capable of detecting biowarfare agents. However, the 2007 NIAID Strategic Plan for Biodefense indicates the need to migrate from the "one bug, one solution" approach, to methods capable of responding to multiple threat organisms. For example, there is a need to identify more than a dozen biowarfare agents and distinguish them from common pathogens that produce similar life threatening critical illness. Another need is for highly adaptable methods, which allow rapid modification as new biothreat agents emerge. ITI is meeting these needs by developing a new system called the FilmArray(tm), which comprises a real-time polymerase chain reaction (PCR) instrument and novel lab-in-a-pouch that performs all steps from sample purification to multiplex organism detection and identification in under 1 hour. A single human sample is injected into the pouch, then inserted in the instrument which automatically performs all required steps. This system is highly adaptable, as pouches can be manufactured with different multiplex assays. Thus far we have developed a pouch that can identify 20 respiratory pathogens, in a 120-well array. In this proposal, ITI will extend FilmArray's capabilities by developing a system that identifies 32 organisms. To do so the following aims will be pursued. Aim 1. Produce FilmArray biothreat assays that identify both biowarfare agents and common pathogens. Aim 2. Increase the number of organisms detected by building a 224-well 2nd stage PCR array. Aim 3. Develop methods for FilmArray automated isolation of bacterial and viral DMA and RNA from nasal swabs and cerebral spinal fluid (CSF). Aim 4. Develop software to facilitate the automated identification of pathogen(s) within a specimen. Aim 5. Validate the FilmArray biothreat system with human samples, including nasal swabs, blood, and CSF at ITI, external laboratories and hospitals. RELEVANCE (See instructions^: The FilmArray will fill a need for rapid, sensitive, specific, easy to use, adaptable, and cost-effective medical diagnostics for public health and hospital-based clinical laboratories, and point-of-care use. FilmArray's automated and simultaneous detection of multiple pathogens will distinguish individuals infected by a biothreat agent or a common infection with similar, generalized symptoms.

描述(由申请人提供)： 针对美国的蓄意恐怖主义行为表明，必须使我国具备抵御这些行动的手段。2001年的炭疽病信件对人类健康造成了严重影响，导致11人住院，5人死亡，以及由于无法快速诊断而产生的恐惧。为了应对这些攻击，爱达荷州科技公司(ITI)创建了能够检测生物制剂的诊断系统。然而，2007年NIAID的生物防御战略计划指出，需要从“一个虫子，一个解决方案”的方法过渡到能够应对多种威胁生物的方法。例如，需要确定十几种生物制剂，并将它们与产生类似威胁生命的危重疾病的常见病原体区分开来。另一个需求是高度适应性的方法，这种方法允许随着新的生物制剂的出现而快速修改。ITI正在通过开发一种名为FilmArray(Tm)的新系统来满足这些需求，该系统包括一个实时聚合酶链式反应(PCR)仪器和新型的袋中实验室，它在不到1小时的时间内完成从样品纯化到多重生物体检测和鉴定的所有步骤。将单个人体样本注入袋中，然后插入仪器，仪器自动执行所有必要的步骤。该系统具有很强的适应性，因为可以用不同的多重检测方法制造袋子。到目前为止，我们已经开发出一种可以识别120孔阵列中的20种呼吸道病原体的袋子。在这项提案中，ITI将通过开发一种识别32种生物的系统来扩展FilmArray的能力。要做到这一点，将追求以下目标。目的1.制作既能鉴定生物制剂又能鉴定常见病原体的膜阵列生物制剂。目的2.建立224孔二级聚合酶链式反应芯片，以增加细菌的检出数量。目的3.建立从鼻拭子和脑脊液中自动分离细菌和病毒DNA和RNA的方法。目的4.开发软件，便于标本中病原体(S)的自动鉴定。目的5.在ITI、外部实验室和医院使用包括鼻拭子、血液和脑脊液在内的人体样本验证FilmArray Biothreat系统。相关性(请参阅说明^：FilmArray将满足公共卫生和医院临床实验室以及医疗保健点使用对快速、灵敏、特定、易于使用、适应性强且经济实惠的医疗诊断的需求。FilmArray对多种病原体的自动同时检测将区分被生物治疗剂感染的个体或具有类似全身症状的普通感染个体。