Bacterial Drug Susceptibility Identification by Surface Enhanced Raman Microscopy

表面增强拉曼显微镜鉴定细菌药敏

• 批准号: 7988796
• 负责人: Alexis F Sauer-Budge
• 金额: $ 91.59万
• 依托单位: FRAUNHOFER CENTER /MANUFACTURING INNOV
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7988796
• 关键词: Accident and Emergency department; Address; Affect; Algorithms; Animals; Antibiotic Resistance; Antibiotics; Antimicrobial Resistance; Bacteremia; Bacteria; Bacterial Infections; Biomedical Engineering; Biophysics; Blood; Blood donor; Cerebrospinal Fluid; Chemistry; Clinical; Clinical Microbiology; Clinical Research; Communicable Diseases; Culture Media; Data; Deposition; Detection; Diagnosis; Diagnostic; Diagnostic tests; Drug resistance; Engineering; Generations; Genetic; Genus staphylococcus; Goals; Gold; Grant; Growth; Health Care Costs; Hour; Human; In Vitro; Infection; Infectious Agent; Label; Laboratories; Libraries; Methicillin Resistance; Microbe; Microscope; Microscopy; Minor; Modeling; Molecular; Morbidity - disease rate; Mus; Nutrient; Optics; Organism; Parasites; Patient Care; Patients; Pharmaceutical Preparations; Pharmacotherapy; Physicians; Point-of-Care Systems; Predisposition; Preparation; Process; Raman Spectrum Analysis; Rattus; Rodent; Sampling; Scientist; Sepsis; Signal Transduction; Simulate; Specificity; Specimen; Spectrum Analysis; Surface; Swab; Symptoms; System; Technology; Testing; Time; United States National Institutes of Health; Urine; antimicrobial; antimicrobial drug; base; communicable disease diagnosis; design; drug resistant bacteria; human subject; improved; meetings; microbial; microorganism; mortality; mutant; nanostructured; new technology; next generation; novel; pathogen; point of care; point-of-care diagnostics; prospective; prototype; response; therapeutic development

DESCRIPTION (provided by applicant): When a patient arrives in an emergency room with clinical symptoms consistent with bloodstream infection, blood cultures are drawn and empiric antimicrobial therapy is given; the actual identification of the pathogen by the laboratory typically takes one or more days. In the absence of specific data on the identity and susceptibility of the pathogen at the time of presentation, the clinician is forced to choose broad-spectrum antimicrobial therapy to cover all possible causes of the suspected bloodstream infection. Unfortunately, such empiric choices can sometimes end up being either ineffective (in the setting of antimicrobial resistance) or unnecessarily broad (in the setting of a susceptible and easily treated organism), potentially increasing morbidity, mortality, and resultant health care costs To address this need, we have developed a prototype identification system based on surface enhanced Raman spectroscopy (SERS). The detection technology consists of a portable Raman microscope, a novel nanostructured substrate, and detection algorithms that have exquisite analytical sensitivity (down to a single bacterium) and specificity (down to the strain level, with the ability to distinguish drug resistant bacteria). Moreover, detection is ultra-fast (~20 sec). To enable this technology to be used at point of care, we have developed an initial prototype system for isolating and concentrating low numbers of bacteria from blood and depositing those bacteria onto the SERS substrate within ~20 min. Here, we propose to build a next generation sample preparation prototype that will be integrated with our existing portable Raman microscope. Furthermore, we will increase the library of Raman signatures to include the most common causes of bacteremia and study the molecular basis for the signatures. The final system will be optimized and validated by testing with samples from human blood cultures and blood obtained directly from animals with experimental bacteremia. To meet these goals, we have assembled a multi-disciplinary team of engineers, basic scientists, and clinician-scientists. At the conclusion of this five-year project, we will have a hardened and tested system which will be ready for clinical studies to diagnose bacteremia in humans. The proposed system will enable identification of microbial pathogens rapidly enough to inform initial antimicrobial drug therapy, thereby reducing morbidity, mortality, and, thereby, healthcare costs. Moreover, the system can be used to address other types of infections by implementing minor changes to the sample processing system to handle additional sample types. We believe that the complete system will revolutionize the field of clinical microbiology by providing a new technology for identifying bacteria and providing basic susceptibility information in time for initial antimicrobial therapy. The system we propose to develop further with this grant will impact the diagnosis and treatment of bacterial infections by enabling clinicians to identify the infectious agent and its antibiotic resistance within half an hour. The physician can then use this information to correctly prescribe a narrow-spectrum antibiotic, which will result in improved patient care and reduced healthcare costs.

描述(申请人提供)：当患者到达急诊室时，临床症状与血流感染一致，抽取血液培养并进行经验性抗菌治疗；实验室对病原体的实际鉴定通常需要一天或几天。在缺乏关于病原体的身份和敏感性的具体数据的情况下，临床医生被迫选择广谱抗菌治疗，以涵盖疑似血流感染的所有可能原因。不幸的是，这种经验性的选择有时可能最终要么无效(在设置抗菌素耐药性方面)，要么不必要地广泛(在设置敏感和容易治疗的生物体的情况下)，潜在地增加发病率、死亡率和由此产生的医疗成本。为了满足这种需求，我们开发了一种基于表面增强拉曼光谱(SERS)的原型识别系统。该检测技术由便携式拉曼显微镜、新型纳米结构底物和具有精致分析灵敏度(低至单个细菌)和特异性(低至菌株水平，能够区分抗药性细菌)的检测算法组成。此外，检测速度极快(约20秒)。为了使这项技术能够在护理点使用，我们开发了一个初步的原型系统，用于从血液中分离和浓缩少量细菌，并在约20分钟内将这些细菌沉积到SERS底物上。在这里，我们建议建立下一代样品制备原型，它将与我们现有的便携式拉曼显微镜集成在一起。此外，我们将增加拉曼特征标记库，以包括最常见的菌血症原因，并研究特征的分子基础。最终的系统将通过测试来自人类血液培养的样本和直接从患有实验性菌血症的动物身上获得的血液进行优化和验证。为了实现这些目标，我们组建了一个由工程师、基础科学家和临床科学家组成的多学科团队。在这个为期五年的项目结束时，我们将拥有一个经过强化和测试的系统，该系统将准备好用于临床研究，以诊断人类的菌血症。拟议的系统将能够足够快地识别微生物病原体，以便为最初的抗菌药物治疗提供信息，从而降低发病率和死亡率，从而降低医疗成本。此外，该系统还可以通过对样本处理系统进行微小更改来处理其他类型的样本，从而解决其他类型的感染问题。我们相信，这个完整的系统将为临床微生物学领域带来革命性的变化，它将提供一种识别细菌的新技术，并为最初的抗菌治疗及时提供基本的药敏信息。我们建议用这笔赠款进一步开发的系统将使临床医生能够在半小时内确定感染源及其抗生素耐药性，从而影响细菌感染的诊断和治疗。然后，医生可以使用这些信息来正确地开出窄谱抗生素，这将导致改善患者护理并降低医疗成本。