FilmArray Direct: Rapid Diagnosis of Antimicrobial-Resistant Pathogens from Blood

FilmArray Direct：快速诊断血液中的耐药病原体

• 批准号: 8875900
• 负责人: Andrew Hemmert
• 金额: $ 140.02万
• 依托单位: BIOFIRE DIAGNOSTICS, LLC
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-03-01 至 2020-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8875900
• 关键词: Adult; Antibiotic Resistance; Antibiotic-resistant organism; Antimicrobial Resistance; Bacteremia; Bacteria; Bacterial DNA; Bacterial Drug Resistance; Base Sequence; Biological Assay; Blood; Blood Volume; Blood specimen; Centrifugation; Chemistry; Childhood; Clinical; Clinical Microbiology; Clinical Sensitivity; Cloaca Chamber; Cyclophosphamide; Cytolysis; DNA amplification; Data; Detection; Diagnosis; Diagnostic; Diagnostic tests; Disadvantaged; Enterobacteriaceae; Enterococcus; Environment; Escherichia coli; Etiology; Failure; Genetic Determinism; Genomic DNA; Gold; Gram-Negative Bacteria; Gram-Positive Bacteria; Hospitals; Hour; Infection; Klebsiella pneumonia bacterium; Laboratories; Life; Medical; Methods; Molecular; Morbidity - disease rate; Nosocomial Infections; Nucleic Acid Amplification Tests; Nucleic acid sequencing; Organism; Outcome; Patient Care; Patients; Population Heterogeneity; Predisposition; Procedures; Production; Proteus; Protocols documentation; Pseudomonas aeruginosa; Reagent; Recovery; Resistance; Resistance profile; Running; Sampling; Sensitivity and Specificity; Sepsis; Staphylococcus aureus; Streptococcus; System; Technology; Testing; Time; Whole Blood; Work; base; clinical practice; diagnosis standard; genetic selection; improved; meetings; microorganism; mortality; novel; pathogen; preclinical evaluation; public health relevance; rapid diagnosis; resistance gene; whole blood bacteria

 DESCRIPTION (provided by applicant): Sepsis is a life-threatening bloodstream infection that results in significant morbidity and mortality. Prompt and appropriate therapy is critical to improved outcomes, but determining the infectious micro-organism can take days. Choice of therapy is further complicated by the dramatic rise in the number of infections caused by antibiotic-resistant organisms. Rapid identification of the etiological agent along with targeted resistance information would enable optimized treatment and improved patient care. The FilmArray(tm) (FA) platform developed and manufactured by BioFire Diagnostics, LLC, is well-suited to meet the challenges of rapid sepsis diagnosis. It is a highly multiplexed, easy to use, sample to result PCR- based diagnostic system. In this project, we propose to develop and optimize a direct from blood FA panel to detect important nosocomial bacteria and their principal genetic determinants of antibiotic resistance. We will incorporate key additional technologies for optimal sensitivity and specificity. This project involves four Aims: Specific Aim 1: Develop and Optimize the FA Sepsis Direct Panel We will develop and optimize a panel for the FA system to simultaneously detect bacteria causing nosocomial infection, as well their primary determinants of antibiotic resistance. This panel will target at least 12 different Gram-negative and Gram-positive bacteria as well as 16 antibiotic resistance genes. Specific Aim 2: Develop External Enrichment Procedure of Bacteria from Blood We will develop and optimize a selective lysis and centrifugation protocol for the concentration of bacterial pathogens from large volumes of blood. With this protocol we will concentrate the viable bacteria present in up to 10 mL of whole blood to a 0.1 mL volume that can be completely introduced into a FA pouch for analysis. Specific Aim 3: Reduce Impact of Bacteria Genomic DNA (bgDNA) Background in the FA pouch We will reduce the impact of bgDNA in FA pouches by 1) working with our raw materials suppliers to lower the amount of bgDNA entering the FA pouch production line and 2) mature technology to modify sample nucleic acid sequences to create non-canonical sequences that can be recognized by unique PCR primers, effectively nullifying the impact of bgDNA. Specific Aim 4: Preclinical Evaluation of the FilmArray Sepsis Direct System in Whole Blood We use the assay system developed in specific aims 1 through 3 to test 1,000 whole blood samples from pediatric and adult patients with suspected sepsis in a preclinical evaluation. We will compare the results of FA Sepsis Direct panel testing to clinical and laboratory data for each patient.

 描述（由申请人提供）：败血症是一种危及生命的血流感染，可导致显着的发病率和死亡率。及时和适当的治疗对于改善结果至关重要，但确定传染性微生物可能需要数天时间。由于抗生素耐药微生物引起的感染数量急剧增加，治疗的选择变得更加复杂。快速识别病原体以及有针对性的耐药信息将有助于优化治疗并改善患者护理。 BioFire Diagnostics, LLC 开发和制造的 FilmArray(tm) (FA) 平台非常适合应对脓毒症快速诊断的挑战。它是一种高度多重、易于使用、从样本到结果的基于 PCR 的诊断系统。在这个项目中，我们建议开发和优化直接来自血液的 FA 检测组，以检测重要的医院细菌及其抗生素耐药性的主要遗传决定因素。我们将采用关键的附加技术来实现最佳的灵敏度和特异性。该项目涉及四个目标： 具体目标 1：开发和优化 FA Sepsis Direct Panel 我们将为 FA 系统开发和优化一个面板，以同时检测引起院内感染的细菌及其抗生素耐药性的主要决定因素。该小组将针对至少 12 种不同的革兰氏阴性和革兰氏阳性细菌以及 16 种抗生素抗性基因。 具体目标 2：开发血液中细菌的外部富集程序 我们将开发和优化选择性裂解和离心方案，用于浓缩大量血液中的细菌病原体。通过该方案，我们将最多 10 mL 全血中存在的活细菌浓缩至 0.1 mL 体积，可以完全引入 FA 袋中进行分析。 具体目标 3：减少 FA 袋中细菌基因组 DNA (bgDNA) 的影响 我们将通过以下方式减少 FA 袋中 bgDNA 的影响：1) 与我们的原材料供应商合作，降低进入 FA 袋生产线的 bgDNA 量；2) 利用成熟的技术修改样品核酸序列，以创建可被独特 PCR 引物识别的非规范序列，从而有效消除 基因组DNA。 具体目标 4：全血中 FilmArray 脓毒症直接系统的临床前评估 我们使用在具体目标 1 至 3 中开发的测定系统，在临床前评估中测试来自疑似脓毒症儿童和成人患者的 1,000 份全血样本。我们会将 FA Sepsis Direct 小组测试的结果与每位患者的临床和实验室数据进行比较。