Rapid, Highly Multiplexed and Sensitive Blood Test for Drug-resistant Bacteria

快速、高度多重且灵敏的耐药细菌血液检测

• 批准号: 7749101
• 负责人: Taiho Kim
• 金额: $ 30万
• 依托单位: NESHER TECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7749101
• 关键词: Address; Affect; Affinity; Antibiotic Resistance; Antibiotics; Antibodies; Applications Grants; Archives; Area; Bacteria; Bacterial Infections; Binding; Biophysics; Blood; Blood Tests; Body Fluids; Botulinum Toxin Type A; Cessation of life; Clinical; Color; Communicable Diseases; Community-Acquired Infections; Complement; Complex; DNA; Detection; Development; Diagnosis; Diagnostic; Diagnostic tests; Diffuse; Discrimination; Drug resistance; Dyes; Energy Transfer; Enterococcus faecalis; Event; Exclusion; Expenditure; Fluorescence; Fluorescence Spectroscopy; Funding; Future; Genetic; Genetic Markers; Goals; Head; Health; Health Care Costs; Healthcare; Hospitalization; Hospitals; Human; Immobilization; Individual; Infection; Infection Control; Infectious Agent; Institution; Intellectual Property; Label; Lasers; Lead; Leadership; Licensing; Life; Measurement; Medical; Medical center; Methicillin; Methodology; Methods; Molecular; Noise; Nucleic Acids; Organism; Patients; Pattern; Phase; Physicians; Procedures; Proteins; RNA; Reader; Reagent; Research; Resistance; Resolution; Sampling; Screening procedure; Serological; Shewanella; Signal Transduction; Small Business Innovation Research Grant; Solutions; Sorting - Cell Movement; Source; Staphylococcus aureus; Symptoms; System; Targeted Toxins; Techniques; Technology; Testing; Therapeutic Intervention; Time; Toxin; United States; Vancomycin; Vancomycin-resistant S. aureus; Virulence; Virulence Factors; antimicrobial; base; cost; drug resistant bacteria; improved; interest; methicillin resistant Staphylococcus aureus; method development; microbial; pathogen; public health relevance; single molecule; therapeutic target; tool; trait; trend; vancomycin B; virtual

DESCRIPTION (provided by applicant): There is an urgent need for rapid, highly sensitive, specific, easy to use, and cost-effective medical diagnostic tests to diagnose individuals exposed to and/or infected with healthcare-associated pathogens that are pre-symptomatic, symptomatic, or have non-specific symptoms, and to accurately identify infectious agents or toxins in clinical samples so appropriate therapeutic intervention can be executed. As antibiotic resistance is spreading rapidly throughout the world causing growing costs in terms of human lives and healthcare expenditures, concomitant determination of increasingly complex microbial resistance patterns is of critical importance. However, as patient samples are limited and pathogen concentrations often extremely low (<10 organisms/ml), serial analysis is not practical, thus requiring highly multiplexed systems instead of individual pathogen-specific tests. Nesher Technologies, Inc.'s (NTI) long-term goal is to develop a rapid, highly multiplexed (with a capacity of >100 analytes per standard patient sample), ultrasensitive and -specific, quantitative, cost-effective, and fully automated, nucleic acid- and protein-based diagnostic system for bacterial infections, to allow quick and accurate pathogen identification in clinical samples, including acquired genetic traits such as antibiotic resistance or enhanced virulence. NTI has licensed the intellectual property for a revolutionary ultrasensitive biodetection technology with exquisite single well multiplexing potential, which was developed at the UCLA Single Molecule Biophysics Lab (headed by Prof. Shimon Weiss). It is based on 3- color alternating laser excitation (3c-ALEX) single molecule fluorescence spectroscopy, whereby two (or three) recognition molecules are tagged with different color fluorescence dyes. Coincident confocal detection of two or three colors constitutes a positive target detection event, allowing molecular identification of freely diffusing molecules in solution and detection of numerous targets simultaneously. Over the Phase I funding period, we will demonstrate feasibility by developing a test to simultaneously distinguish between 11 bacterial strains which are major pathogens responsible for nosocomial and community-acquired infections. Specific aims are: 1. Separate detection and quantification of eight genetic markers (species-specific for Staphylococcus aureus, S. epidermidis, Enterococcus faecalis, E. faecium, and Shewanella oneidensis as positive control; resistance-specific for methicillin, vancomycin A, and vancomycin B) in purified DNA derived from MSSA, MRSA, VRSA, MSSE, MRSE, VSEfaecalis, vanA VREfaecalis, vanB VREfaecalis, VSEfaecium, vanA VREfaecium, vanB VREfaecium, and S. oneidensis by 3-color ALEX-based fluorescence spectroscopy. 2. Multiplexed (single-well) discrimination and quantification of bacteria spiked into human blood. 3. ALEX-based analysis of 350 archived clinical isolates from UCLA's Medical Center (including 25 positive for each of the 11 pathogen types, except for rare VRSA) and statistical analysis. PUBLIC HEALTH RELEVANCE: Nesher Technologies, Inc.'s proposed development of a highly multiplexed, ultrasensitive, quantitative, low- cost automated medical diagnostic test system, capable of quickly identifying specific healthcare-associated bacterial strains and drug resistance patterns among the multitude of possibilities from a single patient sample, radically pushes the limits of current technologies. The improved diagnostic information will enable physicians to better prescribe the appropriate antibiotics to patients and enable hospitals to implement infection control procedures to lower their infection rates, thereby saving human lives and reducing healthcare costs. This will allow limiting the use of broad-spectrum antibiotics, encourage the development of narrowly targeted therapeutics, and help curtailing the marked global trend towards increasing antibiotic drug resistance which is a major concern for treatment and management of infectious diseases, affecting the health of millions of people in the United States and around the world.

描述(由申请人提供)：迫切需要快速、高度敏感、特定、易于使用和成本效益高的医学诊断测试，以诊断暴露于和/或感染了与医疗保健相关的病原体的个人，这些病原体是有症状前期、有症状或没有特定症状的，并准确地识别临床样本中的感染性病原体或毒素，以便进行适当的治疗干预。由于抗生素耐药性正在世界各地迅速传播，导致人类生命和医疗保健支出不断增加，因此确定日益复杂的微生物耐药性模式至关重要。然而，由于患者样本有限，病原体浓度往往极低(&lt；10个微生物/毫升)，连续分析是不现实的，因此需要高度多重的系统，而不是单独的病原体特定测试。奈舍科技有限公司S的长期目标是开发一种快速、高度多元化(每个标准患者样本可检测100个分析物)、超灵敏、特异、定量、经济实惠、基于核酸和蛋白质的全自动化细菌感染诊断系统，以便快速准确地识别临床样本中的病原体，包括获得性遗传特征，如抗生素耐药性或增强的毒力。NTI已为加州大学洛杉矶分校(UCLA)单分子生物物理实验室(由Shimon Weiss教授领导)开发的具有精致单井多路复用潜力的革命性超灵敏生物检测技术授予知识产权。它基于三色交替激光激发(3c-Alex)单分子荧光光谱，其中两个(或三个)识别分子被标记上不同颜色的荧光染料。两种或三种颜色的重合共焦检测构成了一个积极的目标检测事件，允许对溶液中自由扩散的分子进行分子识别，并同时检测多个目标。在第一阶段的资助期内，我们会发展一项测试，以同时区分11种主要病原体，以证明其可行性。这11种细菌是导致院内感染和社区获得性感染的主要病原体。具体目标是：1.用三色荧光光谱法分别检测和定量从MSSA、MRSA、VRSA、MSSE、MRSE、VSEfaecalis、Vana VREfaecalis、VBVREfaecalis、VANA VREfaecalis、VANA VREFecalis、VANA VREfaecalis、VANA VREFecalis、VSE Fececium、VANA VREFfaecum、VANA VREFfaecum、VANA VREFemecum、VANA VREFecicium、VANA VREFececum、VANA VREFecicium、Vana VREFececum、Vana VREFececum、VANA VREFemecum、Vana VREFececum、VANA VREFecicium、Vana VREFecicium、Vana VREFececum、VANA VREFececum、Vana VREFececum、Vana VREFececium、Vana VREFececum、Vana VREFemecum、Vana VREFemecum、Vana VREFecicium2.多重(单孔)鉴别和定量人体血液中的细菌。3.对加州大学洛杉矶分校医学中心350株临床分离株进行了ALEX分析(除罕见的VRSA外，11种病原菌各有25株阳性)，并进行了统计分析。与公共卫生相关：内舍尔科技公司的S建议开发一种高度多元化、超灵敏、定量、低成本的自动化医疗诊断测试系统，能够从单一患者样本的众多可能性中快速识别特定的医疗保健相关细菌菌株和耐药性模式，从根本上突破了当前技术的极限。改进后的诊断信息将使医生能够更好地为患者开出适当的抗生素，并使医院能够实施感染控制程序，以降低他们的感染率，从而拯救人类生命，降低医疗成本。这将允许限制广谱抗生素的使用，鼓励开发有针对性的治疗方法，并有助于遏制抗生素耐药性增加的显著全球趋势。抗生素耐药性是传染病治疗和管理的一个主要问题，影响到美国和世界各地数百万人的健康。