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

• 批准号: 8655138
• 负责人: Vincent Jen-Jr Gau
• 金额: $ 98.72万
• 依托单位: GENEFLUIDICS, INC.
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-15 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8655138
• 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

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.

描述（由申请人提供）：导致许多常见人类感染性疾病（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擅长获得抗菌耐药性的机制。医疗保健提供者对经验性抗生素使用的广泛行为和食物链中抗生素的浸润具有加速和传播耐药性病原体。结果，临床医生的治疗选择较少，尤其是在最有需要的患者中。一个问题的一个例子是甲氧苄啶磺胺甲恶唑（SXT）耐药大肠杆菌的迅速出现，该大肠杆菌在社区环境中占尿道的85-90％。在1990年代之前，β-内酰胺（例如氨苄青霉素（AMP））是急性简单UTI的标准抗菌方案，但是当大肠杆菌对β-内酰胺的耐药性超过25％时被SXT取代。然而，随着使用的增加，SXT耐药性大大增加，喹诺酮类（例如环丙沙星（CIP））成为首选的抗生素。毫不奇怪，耐喹诺酮类药的尿道病正在上升。在MDR病原体有更大问题的医院中，在某些情况下，尿道病大肠杆菌的喹诺酮抗性率现在超过50％。该阶段II NIAID先进技术SBIR应用的目的是开发和验证RAST（快速抗菌敏感性测试），这是一种综合诊断紧凑型系统，使临床医生能够直接护理（POC），基于证据的抗生素选择以治疗急性细菌感染。 RAST通过提供快速（90分钟与2天）和分散的（POC与实验室）测试来解决标准表型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，这是一种用于护理点病原体鉴定（U01 AI082457）的综合诊断生物芯片，用于使用与微纤维学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，我们就完成了几个关键里程碑：（1）3.5小时的基准式RAST协议的开发和临床验证，表现出94％的精度； （2）RAST与临床尿液样品的兼容性，而无需初始细菌分离； （3） 片上电动细菌浓度和测定增强的可行性； （4）使用微通道上的片上细菌培养； （5）用于病原体鉴定的集成微流体弹药筒； （6）基于墨盒的Rast的初步可行性。在当前II期项目中，我们提出了三个特定目的：特定目标1。电动（EK）处理模块的优化和验证，用于减少体积和原位测定增强。目标1的目的是开发一个EK体积减少模块，以在10分钟内富集100倍的样品，并开发一种原位EK增强技术，以提高电化学测定的检测灵敏度10倍。具体目的2。开发Rast弹药筒进行快速表型抗菌敏感性测试。目标2的目的是开发Rast弹药筒和读取器/歧管系统的过程流量和制造过程，包括样品负载，减少EK量，在含有不同兴趣的抗生素的选择性培养基中培养片上样品，以及通过对细菌16S RRNA的定量测量进行定量测量的表型AST。特定目的3。尿液中Rast弹药筒的临床翻译。 AIM 3的目的是对Rast墨盒和读取器/歧管系统进行分析验证，并使用来自怀疑患有UTI的患者的30个未知临床样本进行临床可行性研究。 RAST的开发和验证将遵守医疗设备的质量管理标准（ISO 13485）的建议标准，并遵守完全自动化的短期孵化周期抗微生物易感系统的联邦法规（21 CFR 866.1645）（请参阅商业化计划D.1.3）。在此II阶段应用程序中，成功完成我们的里程碑将是FDA 510（k）提交，以证明该系统基本上等同于谓词设备。研究策略末尾包含一个单独的里程碑和时间表部分。