An Integrated Diagnostic System for Rapid Antimicrobial Susceptibility Testing

用于快速抗菌药物敏感性测试的集成诊断系统

基本信息

批准号：
8314171
负责人：
Vincent Jen-Jr Gau
金额：
$ 99.71万
依托单位：
GENEFLUIDICS, INC.
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-03-15 至 2015-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8314171
关键词：
Academia Accident and Emergency department Accounting Acute Address Ampicillin Antibiotic Resistance Antibiotic Therapy Antibiotics Antimicrobial Resistance Antimicrobial susceptibility Arizona Bacterial Infections Biological Assay Biosensor Ciprofloxacin Clinical Communicable Diseases Complement Detection Development Devices Diagnostic Doctor of Medicine Doctor of Philosophy Enteral Enterobacteriaceae Escherichia coli Expenditure Feasibility Studies Food Chain Gastroenteritis Goals Gram-Negative Bacteria Health Personnel Healthcare Healthcare Systems Hospitals Hour Human In Situ Industry Infection Infiltration Klebsiella Laboratories Lead Measurement Medical Device Microfluidics Modification Molecular Diagnosis National Institute of Allergy and Infectious Disease Oral Outpatients Patient Care Patients Pattern Phase Pneumonia Predisposition Preparation Principal Investigator Process Proteus Protocols documentation Quinolones Reader Regimen Regulation Research Resistance Resistance profile Ribosomal RNA Sampling Scientist Selection for Treatments Small Business Innovation Research Grant System Techniques Technology Test Result Testing TimeLine Translations Trimethoprim-Sulfamethoxazole U-Series Cooperative Agreements Universities Urinary tract infection Urine Uropathogen Uropathogenic E. coli Validation Wound Infection antimicrobial base beta-Lactams biochip commercialization community setting evidence base improved interest multidisciplinary pathogen point of care quinolone resistance sensor

项目摘要

DESCRIPTION (provided by applicant): Pathogens responsible for many of the common human infectious diseases such as urinary tract infection (UTI), gastroenteritis, pneumonia, and wound infections have proven to be highly adept in acquiring mechanisms of antimicrobial resistance. Widespread injudicious practice of empiric antibiotic usage by healthcare providers and infiltration of antibiotics in the food chain have accelerated selection and dissemination of resistant pathogens. As a consequence, clinicians have fewer treatment options, particularly in the most needy patients. An example of the problem was the rapid emergence of trimethoprim- sulfamethoxazole (SXT) resistant E. coli, which accounts for 85-90% of the UTIs in the community setting. Prior to the 1990s, beta-lactams such as ampicillin (AMP) were the standard antimicrobial regimen for acute uncomplicated UTIs, but was replaced with SXT when E. coli resistance against beta-lactams surpassed 25%. With increasing use, however, SXT resistance increased substantially and quinolones such as ciprofloxacin (CIP) became the antibiotic of choice. Not surprisingly, quinolone-resistant uropathogens are on the rise. In hospitals where MDR pathogens are of even greater problem, the quinolone-resistance rate for uropathogenic E. coli has now exceeded 50% in some settings. The goal of this Phase II NIAID Advanced Technology SBIR application is to develop and validate RAST (rapid antimicrobial susceptibility testing), an integrated diagnostic compact system to enable clinicians to direct point-of-care (POC), evidence-based selection of antibiotics for treatment of acute bacterial infections. RAST addresses the major limitations of standard phenotypic AST platforms (e.g., bioMerieux Vitek, BD Phoenix) by providing rapid (90 minutes vs. 2 days) and decentralized (POC vs. laboratory-based) testing. RAST complements our ongoing NIAID Cooperative Agreement, An Integrated Diagnostic Biochip for Point of Care Pathogen Identification (U01 AI082457), for rapid molecular diagnosis urinary tract infections (UTI) using electrochemical biosensors integrated with microfluidics. Since Phase I, we have accomplished several critical milestones: (1) development and clinical validation of a 3.5 hour bench-top RAST protocol showing 94% accuracy; (2) compatibility of RAST with clinical urine samples without need for initial bacterial isolation; (3) feasibility of on-chip electrokinetic bacterial concentration and assay enhancement; (4) on-chip bacterial culture using microchannels; (5) integrated microfluidic cartridge for pathogen identification; and (6) preliminary feasibility of cartridge-based RAST. In the current Phase II project, we propose three Specific Aims: Specific Aim 1. Optimization and validation of electrokinetic (EK) processing modules for volume reduction and in situ assay enhancement. The goal of Aim 1 is to develop an EK volume reduction module for enriching the sample 100-fold within 10 min and to develop an in situ EK enhancement technique for improving the detection sensitivity of the electrochemical assay by 10-fold. Specific Aim 2. Development of the RAST cartridge for rapid phenotypic antimicrobial susceptibility testing. The goal of Aim 2 is to develop the process flow and fabrication process for the RAST cartridge and reader/manifold system, including sample loading, EK volume reduction, on-chip sample culturing in selective media containing different antibiotics of interest, and phenotypic AST by quantitative measurement of bacterial 16S rRNA. Specific Aim 3. Clinical translation of RAST cartridge in urine. The goal of Aim 3 is to perform analytical validation of RAST cartridge and reader/manifold system and a clinical feasibility study using 30 unknown clinical samples from patients suspected to have UTI. The development and validation of RAST will adhere to the recommended standards of Quality Management Standard for Medical Devices (ISO 13485) and federal regulations for fully automated short-term incubation cycle antimicrobial susceptibility system (21 CFR 866.1645)(see Commercialization Plan D.1.3). Successful accomplishment of our milestones in this Phase II application will be followed by FDA 510(k) submission to demonstrate the system is substantially equivalent to a predicate device. A separate Milestones and Timeline section is included at the end of the Research Strategy. PUBLIC HEALTH RELEVANCE: Development of a compact platform capable of rapid pathogen ID and AST directly from patient's samples can provide clinicians at the point of care (e.g., outpatient office, emergency department) with evidenced-based information to start patient-specific antimicrobial treatment only when necessary. Rapid susceptibility test results and alterations in the use of antibiotics, even short-term, have been found to favorably impact patient care and the antibiotic resistance profiles.

描述（由申请人提供）：导致许多常见人类传染病（例如尿路感染（UTI）、胃肠炎、肺炎和伤口感染）的病原体已被证明非常容易获得抗菌素耐药性机制。医疗保健提供者普遍不明智地根据经验使用抗生素，以及抗生素在食物链中的渗透，加速了耐药病原体的选择和传播。因此，临床医生的治疗选择较少，特别是对于最有需要的患者。该问题的一个例子是甲氧苄啶-磺胺甲恶唑 (SXT) 耐药性大肠杆菌的迅速出现，它占社区环境中尿路感染的 85-90%。 20 世纪 90 年代之前，氨苄西林 (AMP) 等 β-内酰胺类药物是治疗急性无并发症性尿路感染的标准抗菌疗法，但当大肠杆菌对 β-内酰胺类药物的耐药性超过 25% 时，被 SXT 取代。然而，随着使用的增加，SXT 耐药性大幅增加，环丙沙星 (CIP) 等喹诺酮类药物成为首选抗生素。毫不奇怪，耐喹诺酮类尿路病原体正在增加。在耐多药病原体问题更为严重的医院，尿路致病性大肠杆菌的喹诺酮类耐药率在某些环境中已超过 50%。 NIAID 先进技术 SBIR II 期应用的目标是开发和验证 RAST（快速抗菌药敏感性测试），这是一种集成诊断紧凑系统，使临床医生能够指导床旁护理 (POC)、基于证据的抗生素选择治疗急性细菌感染。 RAST 通过提供快速（90 分钟与 2 天）和分散式（POC 与基于实验室）测试，解决了标准表型 AST 平台（例如 bioMerieux Vitek、BD Phoenix）的主要局限性。 RAST 补充了我们正在进行的 NIAID 合作协议，即用于护理点病原体识别的集成诊断生物芯片 (U01 AI082457)，用于使用与微流体集成的电化学生物传感器快速分子诊断尿路感染 (UTI)。自第一阶段以来，我们已经实现了几个关键的里程碑：(1) 3.5 小时台式 RAST 方案的开发和临床验证显示出 94% 的准确度； (2) RAST与临床尿样相容，无需进行初始细菌分离； (3) 片上动电细菌浓缩和测定增强的可行性； (4)利用微通道进行片上细菌培养； (5) 用于病原体识别的集成微流控盒； (6)基于盒式RAST的初步可行性。在当前的第二阶段项目中，我们提出了三个具体目标：具体目标 1. 优化和验证动电 (EK) 处理模块，以减少体积和增强原位测定。目标 1 的目标是开发 EK 减容模块，在 10 分钟内将样品富集 100 倍，并开发原位 EK 增强技术，将电化学测定的检测灵敏度提高 10 倍。具体目标 2. 开发用于快速表型抗菌药物敏感性测试的 RAST 试剂盒。目标 2 的目标是开发 RAST 试剂盒和读取器/歧管系统的工艺流程和制造工艺，包括样品加载、EK 体积减少、在含有不同目标抗生素的选择性培养基中进行片上样品培养，以及通过以下方式进行表型 AST细菌 16S rRNA 的定量测量。具体目标 3. RAST 试剂盒在尿液中的临床转化。目标 3 的目标是对 RAST 盒和读取器/歧管系统进行分析验证，并使用来自疑似患有 UTI 的患者的 30 个未知临床样本进行临床可行性研究。 RAST 的开发和验证将遵循医疗器械质量管理标准 (ISO 13485) 的推荐标准和全自动短期孵化周期抗菌敏感性系统联邦法规 (21 CFR 866.1645)（参见商业化计划 D.1.3）。在成功完成第二阶段申请中的里程碑之后，我们将提交 FDA 510(k)，以证明该系统基本上等同于谓词设备。研究策略末尾包含单独的里程碑和时间表部分。公共卫生相关性：开发一个能够直接从患者样本中快速识别病原体和 AST 的紧凑平台，可以为护理点（例如门诊、急诊科）的临床医生提供基于证据的信息，以便仅开始针对患者的抗菌治疗必要时。人们发现，快速药敏试验结果和抗生素使用的改变，即使是短期的，也会对患者护理和抗生素耐药性产生有利影响。