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）：脑膜炎和脑炎是中枢神经系统的炎性（通常为感染性）过程，可导致显著的发病率和死亡率。及时，适当的治疗是至关重要的，但确定感染病因可能是困难和耗时的。可能涉及广泛的病原体，包括细菌和病毒。细菌培养是诊断细菌性脑膜脑炎（ME）的标准，病毒核酸PCR是无菌性病因的标准。然而，目前的方法要么1）太慢而不具有临床相关性; 2）技术复杂; 3）靶点数量有限，要么4）未被FDA批准。爱达荷州技术公司（ITI）开发了一种名为“FilmArray“的”袋中实验室”PCR诊断系统，该系统快速、易于使用，可同时检测大量传染性病原体。可检测15种呼吸道病原体的FilmArray测试条被FDA批准为CLIA“中等复杂性”检测。我们建议将FilmArray技术应用于ME诊断问题。SA1：开发脑炎脑膜炎多重病原体核酸联检试剂盒（FAME）：我们将构建一个FilmArray测试条，该测试条将现有测试条的PCR检测试剂与五种新检测试剂相结合，以直接从脑脊液（CSF）中检测以下ME致病微生物：肠杆菌科、大肠埃希菌。coli、Enterococcus species、H.流感病毒，L.单核细胞增多症、肺炎支原体、N.脑膜炎、铜绿假单胞菌、葡萄球菌属、S.金黄色葡萄球菌、链球菌属、S. agalactiae、无乳链球菌S.肺炎链球菌（S.化脓性链球菌、绿色链球菌、腺病毒、肠道病毒、副肠孤病毒、HSV 1、HSV 2、VZV、EBV、CMV、HHV-6。SA2：优化CSF中病毒和细菌的核酸提取：CSF是在FilmArray上处理的复杂性较低的样本基质之一（与鼻拭子、血培养和粪便相比）。然而，直接从CSF中检测细菌和病毒可能需要最高的灵敏度。我们将优化测试条内部的核酸纯化步骤，并研究在测试条SA 3之前浓缩细菌和病毒的简单步骤：在300份CSF样本上测试FAME试剂盒：我们的合作者将测试疑似ME的儿童和成人患者的CSF样本。我们将这些结果与他们对样本进行的标准检测进行比较，补充了SA 1中所述的对照试验的结果。该提案的成功结果将是能够检测和识别常见ME病原体的FilmArray测试条。它将为向FDA提交510（k）所需的分析和临床试验做好准备。这将是今后第二阶段划界案的主题。