Rapid, automated, detection of viral and bacterial pathogens causing meningitis

快速、自动化检测引起脑膜炎的病毒和细菌病原体

• 批准号: 8453904
• 负责人: Andrew Hemmert
• 金额: $ 30万
• 依托单位: BIOFIRE DEFENSE, LLC.
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-01-01 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8453904
• 关键词: Acute; Adenoviruses; Adult; Affect; Bacteria; Biological; Biological Assay; Blood; Buffers; CNS processing; Caring; Cerebrospinal Fluid; Childhood; Clinical; Clinical Sensitivity; Clinical Trials; Complex; Culture Media; Cytomegalovirus; DNA-Directed DNA Polymerase; Detection; Development; Diagnosis; Diagnostic; Encephalitis; Enterobacteriaceae; Enterococcus; Enterovirus; Escherichia coli; Etiology; Evaluation; Feces; Future; Genus staphylococcus; Gold; Haemophilus influenzae; Herpesvirus 1; Hour; Housing; Human Herpesvirus 2; Human Herpesvirus 4; Human Herpesvirus 6; Idaho; In Vitro; Infection; Infectious Agent; Inflammatory; Laboratories; Life; Listeria monocytogenes; Medical; Meningitis; Meningoencephalitis; Methods; Morbidity - disease rate; Mycoplasma pneumoniae; Neisseria meningitidis; Nose; Nucleic Acid Amplification Tests; Nucleic Acids; Organism; Outcome; Parechovirus; Pathogen detection; Patients; Performance; Phase; Polymerase Chain Reaction; Population; Process; Protocols documentation; Pseudomonas aeruginosa; Publishing; Resources; Sampling; Sensitivity and Specificity; Soil; Spinal Cord; Staphylococcus aureus; Streptococcus; Streptococcus Group B; Streptococcus pneumoniae; Streptococcus pyogenes; Swab; System; Technology; Testing; Time; United States Food and Drug Administration; Viral; Virus; base; clinical practice; clinically relevant; diagnosis standard; effective therapy; mortality; new technology; nucleic acid purification; pathogen; preclinical evaluation; public health relevance; rapid detection; respiratory; skills; viral DNA; viral detection

DESCRIPTION (provided by applicant): Meningitis and encephalitis are inflammatory, often infectious, processes of the central nervous system that can result in significant morbidity and mortality. Prompt, appropriate therapy is crucial, but determining the infectious etiology can be difficult and time-consuming. A wide-range of pathogens can be involved including both bacteria and viruses. Culture is the standard for diagnosis of bacterial Meningoencephalitis (ME), PCR of viral nucleic acid is the standard for "aseptic" causes. However the current methods are either 1) too slow to be clinically relevant; 2) technically complex; 3) limited in the number of targets r 4) not cleared by the FDA. Idaho Technology (ITI) has developed a "lab in a pouch" PCR-based diagnostic system termed "FilmArray" that is rapid, easy to use, and can test for large panels of infectious agents simultaneously. A FilmArray pouch that can detect 15 respiratory pathogens was cleared by the FDA as a CLIA "moderate complexity" test. We propose to apply the FilmArray technology to the problem of ME diagnosis. SA1: Develop a FilmArray Meningoencephalitis (FAME) panel: We will construct a FilmArray pouch that combines PCR assays from existing panels with five new assays to detect the following ME-causing organisms directly from cerebrospinal fluid (CSF): Enterobacteriaceae, E. coli, Enterococcus species, H. influenza, L. monocytogenes, Mycoplasma pneumoniae, N. meningitidis, P. aeruginosa, Staphylococci species, S. aureus, Streptococcus species, S. agalactiae, S. pneumoniae, S. pyogenes, viridians streptococci, Adenovirus, Enterovirus, Parechovirus, HSV1, HSV2, VZV, EBV, CMV, HHV-6. SA2: Optimize nucleic acid extraction for virus and bacteria from CSF: CSF is one of the less complex sample matrices that have been processed on the FilmArray (when compared to nasal swabs, blood culture and stool). However, detecting bacteria, and virus, directly from CSF, may require the highest possible sensitivity. We will optimize the nucleic acid purification steps internal to the pouch as well as investigate simple steps to concentrate bacteria and viruses before the pouch SA3: Test the FAME panel on 300 CSF samples: Our collaborators will test CSF samples from pediatric and adult patients with suspected ME. We will compare these results to their standard assays performed on the samples, supplemented with the results of the comparator assays described in SA1. A successful outcome of this proposal will be a FilmArray pouch capable of detecting and identifying the common ME pathogens. It will be ready for the analytical and clinical trials necessary for a 510(k) submission to the FDA. This would be the subject of a future phase II submission.

描述（由申请人提供）：中枢神经系统的脑膜炎和脑炎是炎症性的，通常是传染性的过程，可能导致明显的发病率和死亡率。迅速，适当的治疗至关重要，但是确定感染性病因可能很困难且耗时。可能涉及大量的病原体，包括细菌和病毒。培养是诊断细菌脑膜脑炎（ME）的标准，病毒核酸的PCR是“无菌”原因的标准。但是，目前的方法要么是1）太慢，无法在临床上相关； 2）技术复杂； 3）限制目标数r 4）未清除FDA。爱达荷州技术（ITI）开发了一个基于PCR的诊断系统中的“实验室”，称为“ FilmArray”，该系统快速，易于使用，可以同时测试大型传染性剂。 FDA清除了一个可以检测到15种呼吸道病原体的胶片袋作为CLIA“中等复杂性”测试。我们建议将电影制品技术应用于我的诊断问题。 SA1：开发胶卷脑膜脑炎（名望）面板：我们将构建一个胶卷袋，将现有面板中的PCR分析与五个新测定结合在一起，以直接从脑脊液（CSF）直接检测到以下ME引起的有机体（CSF）：肠杆菌科，E。coli，coli，coli，coli，coli，coli coli coli，coli coli，uttrocococcus，h.h.throcmant，h.thropmaint，l.thropmaint，l.thropmaint，l.thropmaint，l.thropmaint，l.throply tharcyn s。 pneumoniae, N. meningitidis, P. aeruginosa, Staphylococci species, S. aureus, Streptococcus species, S. agalactiae, S. pneumoniae, S. pyogenes, viridians streptococci, Adenovirus, Enterovirus, Parechovirus, HSV1, HSV2, VZV, EBV, CMV, HHV-6。 SA2：从CSF：CSF优化病毒和细菌的核酸提取是在膜片上处理过的较不复杂的样品矩阵之一（与鼻拭子相比，与鼻拭子，血液培养和粪便相比）。但是，直接从CSF中检测细菌和病毒可能需要最高的敏感性。我们将优化小袋内部内部的核酸纯化步骤，并研究在小袋SA3之前浓缩细菌和病毒的简单步骤：在300 CSF样品上测试成名面板：我们的合作者将测试来自儿科和成人患者的CSF样品。我们将将这些结果与对样品进行的标准测定进行比较，并补充了SA1中描述的比较器测定结果。该提案的成功结果将是一个能够检测和识别常见的ME病原体的电影袋。它将为FDA提交510（k）所需的分析和临床试验做好准备。这将是未来第二阶段提交的主题。