Rapid and Enclosed System for the Identification of Multi-Drug Resistant TB

用于识别多重耐药结核病的快速封闭系统

基本信息

批准号：
7394908
负责人：
SEASON S-S WONG
金额：
$ 14.52万
依托单位：
LYNNTECH, INC.
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-05-15 至 2010-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7394908
关键词：
Academic Medical Centers Acquired Immunodeficiency Syndrome Africa Agreement Air Antibiotics Antitubercular Agents Area Arts Bacillus (bacterium)Biological Assay Blood capillaries Buffers Caliber Caring Cause of Death Cessation of life Characteristics Chemical Engineering China Clinic Clinical Collaborations Collection Communicable Diseases Complex Condition Country Coupled Data Data Display Deposition Detection Developed Countries Developing Countries Development Devices Diagnosis Diagnostic Diagnostic Procedure Diagnostic Services Diagnostic tests Differential Diagnosis Diffusion Dimensions Disease Drug Resistant Tuberculosis Drug resistance Drug resistance in tuberculosis Drug-sensitive Encephalitis Engineering Equipment Evaluation Fluorescence Foundations Glass Goals Government HIV Health Professional Healthcare Healthy People 2010 Hour Housing Human Immune system Immunologic Deficiency Syndromes In Vitro Incidence Income Individual Infection Infectious Diseases Research Investments Kazakhstan Laboratories Lateral Lead Length Licensing Life Liquid substance Malaria Marketing Medical Medicine Methods Microbe Microbiology Microfluidics Mission Molecular Molecular Target Monitor Multi-Drug Resistance Multidrug-Resistant Tuberculosis Mutation Mycobacterium tuberculosis Nature Nucleic Acids Numbers Oligonucleotides Pathology Patients Personal Satisfaction Persons Pharmaceutical Preparations Phase Pliability Polymerase Chain Reaction Polymers Population Predisposition Preparation Procedures Process Production Public Health Pump Purpose Range Rate Reaction Reagent Recording of previous events Reporting Research Personnel Resistance Respiratory Tract Infections Rifampin Risk Sampling Scientist Screening procedure Sensitivity and Specificity Services Solid Solutions Specimen Sputum Stream Surface Surveys System Techniques Technology Technology Transfer Test Result Testing Texas Time Tube Tuberculosis USSR United States Universities University Hospitals Viral Week Work World Health Organization assay development base capillary commercialization concept cost day design disorder control drug standard evaluation/testing experience falls foodborne innovation instrumentation interest isoniazid killings multidisciplinary novel diagnostics nucleic acid detection pandemic disease pathogen prevent professor rapid diagnosis reaction rate research and development statistics success tool tuberculosis drugs tuberculosis treatment

项目摘要

DESCRIPTION (provided by applicant): Tuberculosis (TB) caused by Mycobacterium tuberculosis is the seventh leading cause of death worldwide, second only to human immunodeficiency virus (HIV) and acquired immunodeficiency syndrome (AIDS) among infectious diseases. In 2004, an estimated 8.9 million people developed TB and 1.7 million died. This situation is compounded by the HIV pandemic with almost 13 million people currently co-infected with HIV and TB, and the emergence of multidrug resistant TB (MDR-TB). In the Unites States, there are about 15,000 new cases of TB reported each year, and approximately 10% of them have resistance to drugs. Rapid diagnosis of TB infections and identification of drug resistance is key to disease control and treatment. In order to accomplish the Healthy People 2010 goal of reducing the time required for the laboratory confirmation of the diagnosis of tuberculosis to 48 hours, rapid tests to detect Mycobacterium tuberculosis or its products are needed. In addition, rapid tests that can reduce the turnaround-time for detection of drug- resistance are needed as well. It is clear that no test is yet available that meets target specifications, and new methods that can overcome limitations and respond to the challenges posed will be well received. Our proposed work will employ state-of-the-art technology to provide differential diagnostics to identify Mycobacterium tuberculosis infection and determine if it has mutations that cause resistances to the most commonly used antibiotics for TB. The goal is to achieve sample-to-result testing in less than 90 minutes. After sample preparation using automated nucleic acid (NA) extraction instrumentation, the NA template will be drawn into an in-line ultra-fast PCR reaction where a highly multiplexed amplification assay (covering 25 mutation/targets) will take place. This powerful multiplex amplification technology from our collaborator overcomes the obstacles of traditional multiplex PCR assay development. It allows for specific and sensitive amplification of multiple targets in a single reaction. The PCR products, mixed with hybridization buffer, will then be transported downstream in a closed format to perform an extremely simple microfluidic based multiplexed detection. The PCR product will be passed over a linear glass capillary substrate with spatially addressable capture zones that are modified with different oligonucleotide capture probes. The smaller diameter linear glass substrate will be housed in a slightly larger diameter polymeric tubing to provide an "array-in-a tube" conditions with microfluidic characteristics for rapid and sensitive fluorescence detection utilizing an extremely simple and inexpensive setup. In Phase I, a breadboard system will be developed and the technology feasibilities will be demonstrated with the nucleic acid templates provided by collaborators. Phase I results will enable the subsequent design and development of an automated clinical diagnostic assay that will fulfill a crucial need in TB care, as well as the detection of other diseases. Tuberculosis (TB), which claims nearly 2 million lives each year, is the seventh leading cause of death worldwide, second only to HIV/AIDS among infectious diseases. Making the condition worse, the emergence of multidrug resistant TB (MDR-TB) makes the treatment more difficult and costly. In the Unites States, there are about 15,000 new cases of TB reported each year, and approximately 10% of them have resistance to drugs. A quick and accurate diagnostics tool for TB and MDR-TB is key to disease control and treatment and could save up to 625,000 of those lives each year (Nature S1, 46-57; 2006).

描述（申请人提供）：由结核分枝杆菌引起的结核病（TB）是全球第七大死因，在传染病中仅次于人类免疫缺陷病毒（HIV）和获得性免疫缺陷综合症（AIDS）。 2004年，估计有890万人患上结核病，170万人死亡。由于艾滋病毒大流行（目前有近 1300 万人同时感染艾滋病毒和结核病）以及耐多药结核病（MDR-TB）的出现，情况变得更加复杂。在美国，每年报告约15,000例新发结核病病例，其中约10%具有耐药性。结核病感染的快速诊断和耐药性鉴定是疾病控制和治疗的关键。为了实现“健康人2010”目标，将实验室确认结核病诊断所需的时间减少到48小时，需要快速检测结核分枝杆菌或其产物。此外，还需要能够缩短耐药性检测周转时间的快速测试。显然，目前还没有满足目标规范的测试，能够克服限制并应对所提出的挑战的新方法将受到广泛欢迎。我们提出的工作将采用最先进的技术来提供鉴别诊断，以识别结核分枝杆菌感染，并确定它是否具有导致对最常用的结核病抗生素产生耐药性的突变。目标是在 90 分钟内实现从样品到结果的测试。使用自动核酸 (NA) 提取仪器制备样品后，NA 模板将被引入在线超快速 PCR 反应中，其中将进行高度多重扩增测定（涵盖 25 个突变/靶标）。我们的合作者提供的这种强大的多重扩增技术克服了传统多重 PCR 检测开发的障碍。它允许在单个反应中特异性且灵敏地扩增多个靶标。 PCR 产物与杂交缓冲液混合，然后以封闭形式向下游输送，以执行极其简单的基于微流体的多重检测。 PCR 产物将通过线性玻璃毛细管基板，该基板具有空间可寻址的捕获区，这些捕获区用不同的寡核苷酸捕获探针进行修饰。较小直径的线性玻璃基板将被容纳在稍大直径的聚合物管中，以提供具有微流体特性的“管内阵列”条件，从而利用极其简单和廉价的装置进行快速和灵敏的荧光检测。在第一阶段，将开发面包板系统，并利用合作者提供的核酸模板来证明技术可行性。第一阶段的结果将使自动化临床诊断测定的后续设计和开发成为可能，该测定将满足结核病护理以及其他疾病检测的关键需求。结核病（TB）每年夺去近200万人的生命，是全球第七大死因，在传染病中仅次于艾滋病毒/艾滋病。耐多药结核病（MDR-TB）的出现使病情变得更糟，使治疗变得更加困难和昂贵。在美国，每年报告约15,000例新发结核病病例，其中约10%具有耐药性。结核病和耐多药结核病快速准确的诊断工具是疾病控制和治疗的关键，每年可挽救多达 625,000 人的生命（Nature S1，46-57；2006）。