A fully integrated CentriFluidic system for direct bloodstream infection PID/AST

用于直接血流感染 PID/AST 的完全集成 CentriFluidic 系统

• 批准号: 9241943
• 负责人: Vincent Jen-Jr Gau
• 金额: $ 70.42万
• 依托单位: GENEFLUIDICS, INC.
• 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 2020-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9241943
• 关键词: Acinetobacter baumannii; Acute; Address; Agreement; Algorithmic Analysis; Antibiotic Resistance; Antibiotics; Antimicrobial susceptibility; Automation; Bacteremia; Bacteria; Bacterial Infections; Biological Assay; Blood; Blood Cells; Blood specimen; Blood-Borne Pathogens; Categories; Centrifugation; Characteristics; Clinical; Clinical Management; Clinical Sensitivity; Culture Techniques; Cytolysis; Detection; Devices; Diagnosis; Diagnostic; Dose; Enterobacter; Escherichia coli; Etiology; Feasibility Studies; Genotype; Goals; Gravitation; Guidelines; Hospitals; Hour; Hybrids; Incidence; Infection; Inflammatory Response; Intestines; Klebsiella pneumonia bacterium; Libraries; Liquid substance; Mass Spectrum Analysis; Minimum Inhibitory Concentration measurement; Modality; Modification; Molecular; Molecular Analysis; Monitor; Multi-Drug Resistance; Observational Study; Organism; Patients; Phenotype; Physicians; Predisposition; Procedures; Process; Protocols documentation; Pseudomonas aeruginosa; Public Health; Publishing; Recovery; Regimen; Reporting; Reproducibility; Research; Research Project Grants; Resistance profile; Sampling; Sensitivity and Specificity; Sepsis; Septic Shock; Specificity; Specimen; Survival Rate; System; Systems Integration; Technology; Testing; Time; Ultracentrifugation; Urinary tract infection; Urine; Whole Blood; antimicrobial; base; blind; clinically relevant; evidence base; improved; innovation; melting; microorganism; mortality; pathogen; precision medicine; prospective; prototype; public health relevance; routine Bacterial stain; time use; validation studies; Acinetobacter baumannii; Acute; Address; Agreement; Algorithmic Analysis; Antibiotic Resistance; Antibiotics; Antimicrobial susceptibility; Automation; Bacteremia; Bacteria; Bacterial Infections; Biological Assay; Blood; Blood Cells; Blood specimen; Blood-Borne Pathogens; Categories; Centrifugation; Characteristics; Clinical; Clinical Management; Clinical Sensitivity; Culture Techniques; Cytolysis; Detection; Devices; Diagnosis; Diagnostic; Dose; Enterobacter; Escherichia coli; Etiology; Feasibility Studies; Genotype; Goals; Gravitation; Guidelines; Hospitals; Hour; Hybrids; Incidence; Infection; Inflammatory Response; Intestines; Klebsiella pneumonia bacterium; Libraries; Liquid substance; Mass Spectrum Analysis; Minimum Inhibitory Concentration measurement; Modality; Modification; Molecular; Molecular Analysis; Monitor; Multi-Drug Resistance; Observational Study; Organism; Patients; Phenotype; Physicians; Predisposition; Procedures; Process; Protocols documentation; Pseudomonas aeruginosa; Public Health; Publishing; Recovery; Regimen; Reporting; Reproducibility; Research; Research Project Grants; Resistance profile; Sampling; Sensitivity and Specificity; Sepsis; Septic Shock; Specificity; Specimen; Survival Rate; System; Systems Integration; Technology; Testing; Time; Ultracentrifugation; Urinary tract infection; Urine; Whole Blood; antimicrobial; base; blind; clinically relevant; evidence base; improved; innovation; melting; microorganism; mortality; pathogen; precision medicine; prospective; prototype; public health relevance; routine Bacterial stain; time use; validation studies

 DESCRIPTION (provided by applicant): There is a pressing need for a technology platform that can aid rapid AST profiling where MIC results are presented within a narrow time window of less than 3 hours. This acquires tremendous significance during bloodstream infections and rapid onset sepsis; the choice of antibiotics to surmount emergent antibiotic resistance and associated treatment modality in reduction of host inflammatory response is crucial for patient survival and recovery. Such a platform will finally usher in the "Precision medicine" approach for treating bacterial infections, whereby physicians are able to track antibiotic resistance profiles f pathogens in real time and adjust their antibiotic dosing regimens. It is highly unlikely that culture techniques can still be improved to significantly decrease total assay time (TAT). Therefore, this proposed system focuses on improving TAT by using molecular technologies for identification and characterization of microorganism susceptibility profile directly from fresh whole blood specimen of BSI patients. A first step forward, to decrease time-to-result, can be achieved by utilizing blood lysis centrifugation. Moreover, eliminating steps for obtaining clinica isolates will lead to the most optimal turnaround time. The overall hypotheses of this application are that a CentriFluidic system can be used to perform a rapid PID and AST directly from whole blood without a need of blood cultivation or PCR amplification based on our successful clinical feasibility studies on urinary tract infection (UTI) directly from raw urine specimen with reproducible demonstration of polymicrobial infection detection and multi-drug resistance profiling. We developed, demonstrated and published an innovative molecular-based genotypic-phenotypic-hybrid approach for multiplexed bacterial PID and AST profiling with 100% clinical sensitivity, 96% clinical specificity, 98% minimum inhibitory concentration (MIC) and 97% categorical agreement in our most recent ongoing clinical feasibility study on 73 raw clinical urine samples. However, critical limitations of the current UTI platform exist, including inabilityto detect much lower abundance of bloodborne pathogens, the lack of system integration with an ultracentrifugation module and inability to cover all clinical relevant pathogens for BSIs. The main goal of this RO1 research project is to combine the advantages of lab automation, rapid molecular analysis, melt-curve signature analysis, genotypic pathogen quantification and phenotypic antibiotic conditions to dramatically improve the sensitivity and specificity of rapid, evidence-based PID and AST directly from patient blood samples. We propose to test the following hypotheses: Specific Aim 1: Transition the current electrochemical-based molecular analysis PID/AST platform technology from UTI to BSIs - Hypothesis: Lysis centrifugation can address the change of matrix effect from raw urine to whole blood samples and the issue of low abundant pathogen for BSIs. Specific Aim 2: Develop a dual-mode electrochemical-based dynamic hybridization analysis algorithm to expand the BSI PID/AST panel - Hypothesis: Dynamic hybridization analysis can be utilized to expand the species-specific identification of common and emerging pathogens for BSIs. Specific Aim 3: Prototype, validate and manufacture the CentriFluidic cartridge and system - Hypothesis: Ultracentrifugation (up to 50,000 g, gravitational force) can be incorporated into a multiplexed fluidic cartridge for a fully automated BSIs PID/AST from whole blood samples in 3 hours. Specific Aim 4: Clinically validate the rapid BSI PID/AST CentriFluidic system according to CLSI guidelines - Hypothesis: Blood samples spiked with ATCC strain bacteria used in the analytical validation studies represent critical matrix characteristics of fresh whole blood samples from patients.

 描述（由适用提供）：对于技术平台的迫切需求，该平台可以帮助快速分析，其中在小于3小时的狭窄时间窗口内显示了麦克风结果。这在血液感染和快速发作败血症过程中获得了极大的意义。在减少宿主炎症反应中，选择抗生素以克服新兴抗生素耐药性和相关治疗方式对于患者的生存和康复至关重要。这样的平台最终将引入治疗细菌感染的“精密医学”方法，医生能够实时跟踪抗生素耐药性F病原体并调整其抗生素剂量方案。培养技术仍然很不可能得到改进，以显着减少总测定时间（TAT）。因此，该提出的系统着重于通过使用分子技术直接从BSI患者的新鲜全血标本中鉴定和表征微生物敏感性概况来改善TAT。可以通过使用血液裂解离心来实现缩短时间的第一步。此外，消除获得临床分离株的步骤将导致最佳的周转时间。该应用程序的总体假设 我们开发，演示和发表了创新的基于分子的分析。用于100％临床敏感性，96％的临床特异性，98％最低抑制浓度（MIC）和97％分类一致性的基因型 - 表型杂交方法和AST分析，并具有100％的临床敏感性，96％的临床特异性，97％的分类一致性在我们最近进行的73个原始临床尿液样本上的持续临床可行性研究中。但是，存在当前UTI平台的临界局限性，包括无法检测到较低的血源性病原体，缺乏与超速离心模块的系统整合以及无法涵盖BSIS的所有临床相关病原体。该RO1研究项目的主要目的是结合实验室自动化，快速分子分析，融化曲线签名分析，基因型病原体定量和表型抗生素条件的优势，以显着提高快速，证据的基于证据的PID的敏感性和特异性，直接来自患者血液样本。我们建议检验以下假设：特定目标1：过渡当前基于电化学的分子分析PID/AST平台技术从UTI到BSIS-假设：裂解离心可以解决从原始尿液到全血液样品的基质效应的变化，以及BSIS的低大量病原体问题。具体目的2：开发基于双模式电化学的动态杂交分析算法以扩展BSI PID/AST面板 - 假设：动态杂交分析可用于扩展BSIS的常见和出现病原体的特定规范鉴定。特定目标3：原型，验证和制造离心弹药筒和系统 - 假设：可以将超速离心（高达50,000 g，重力力）掺入多重液体墨盒中，以进行全自动的墨盒 BSIS PID/AST在3小时内从全血样品中获得。具体目标4：根据CLSI指南，在临床上验证快速BSI PID/AST离心系统 - 假设：在分析验证研究中使用的ATCC菌株峰值的血液样本代表了患者新鲜全血样品的关键基质特征。