Miniarray Detection of MTB Complex Species

MTB 复杂物种的微阵列检测

• 批准号: 6742764
• 负责人: DAVID A SHAFER
• 金额: $ 10万
• 依托单位: WEILIN BIOTECHNOLOGY, INC.
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-02-01 至 2005-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6742764
• 关键词: Mycobacterium tuberculosis; bacterial genetics; diagnosis design /evaluation; diagnostic tests; genetic strain; microarray technology; nucleic acid amplification techniques; technology /technique development

DESCRIPTION (provided by applicant): Tuberculosis is the most important cause of death worldwide from a single pathogen. Diagnosis primarily occurs by microscope analysis of stained sputum smears and from growing cultures for biological tests. While staining is quick, it is uncertain, and while culturing is definitive, it is slow. Proprietary tests based on nucleic acid amplification are fast and reliable, but they require equipment and reagents only suited to advanced labs in developed countries. A new miniarray test system is proposed that combines the speed and sensitivity advantages of amplification-based assays but employs a simplified microarray detection platform, called miniarrays, that reduces cost and complexity and that provides discrimination of multiple species in a single test. Features of this miniarray platform also promise to overcome the need for PCR amplification in latter versions of the assay by employing proprietary signal amplification methods and reagents. Thus, two product lines are expected: 1. a high-tech version for developed countries to provide greater diagnostic capability as well as a fast and more reliable front-line test, and 2. a Iow-tech bench-top analysis version for developing countries that enables fast discrimination of multiple species but achieves diagnosis without PCR equipment, high-resolution microarray scanners or fluorescent microscopes. Phase I will focus on developing and testing two prototype miniarrays for discriminating major MTB complex subspecies. While the initial miniarray prototypes remain based on PCR amplification, they will demonstrate visually-simple, species-specific diagnostic patterns reflecting the presence and absence of critical gene segments that characterize a series of related but discrete species or subspecies. Phase II will bring these products to market, extend them to the diagnosis of other important pathogens, and will initiate development of similar Iow-tech miniarrays for developing countries. This technology is based on proprietary, patent-pending reagents and procedures that convert genomic DNA samples into amplified target copies that are modified with universal linkers. These special linkers then enable global amplification of the set of targets with a single primer set, and they also provide means to increase signaling by virtue of binding long DNA-based reporters called GeneTAGs. The associated miniarray platform also offers a path to developing a variety of similar diagnostic biochips for discriminating other pathogens. This miniarray system will be rapid (<2 hrs), sensitive (equal to PCR), inexpensive, and easy to perform. The worldwide market for TB testing is $900M with amplification-based tests constituting approximately 1/3rd of the total. Based on a survey of potential customers and diagnostics professionals, the market potential of our system is over $100M. Successful achievement of developmental milestones could push that potential higher.

描述（由申请人提供）：结核病是全世界单一病原体导致死亡的最重要原因。诊断主要通过染色痰涂片的显微镜分析和用于生物测试的生长培养物进行。虽然染色速度快，但不确定；虽然培养是确定的，但速度慢。基于核酸扩增的专有检测快速可靠，但所需设备和试剂仅适合发达国家的先进实验室。提出了一种新的微型阵列测试系统，该系统结合了基于扩增的测定的速度和灵敏度优势，但采用了称为微型阵列的简化微阵列检测平台，该平台降低了成本和复杂性，并在单次测试中提供了对多个物种的区分。该微型阵列平台的特点还有望通过采用专有的信号放大方法和试剂来克服后面版本的检测中对 PCR 扩增的需求。因此，预计有两条产品线：1.为发达国家提供的高科技版本，提供更强的诊断能力以及快速、更可靠的一线测试；2.为发展中国家提供的低技术台式分析版本，能够快速区分多个物种，但无需PCR设备、高分辨率微阵列扫描仪或荧光显微镜即可实现诊断。第一阶段将重点开发和测试两个原型微型阵列，用于区分主要的 MTB 复杂亚种。虽然最初的微阵列原型仍然基于 PCR 扩增，但它们将展示视觉上简单的物种特异性诊断模式，反映表征一系列相关但离散的物种或亚种的关键基因片段的存在和不存在。第二阶段将把这些产品推向市场，将其扩展到其他重要病原体的诊断，并将启动为发展中国家开发类似的低技术微阵列。 该技术基于正在申请专利的专有试剂和程序，可将基因组 DNA 样本转化为经过通用接头修饰的扩增目标副本。然后，这些特殊的接头能够使用单个引物组对靶标组进行全局扩增，并且它们还提供了通过结合称为 GeneTAG 的基于长 DNA 的报告基因来增强信号传导的方法。相关的微型阵列平台还提供了开发各种类似的诊断生物芯片以区分其他病原体的途径。该微型阵列系统快速（<2 小时）、灵敏（相当于 PCR）、廉价且易于执行。全球结核病检测市场规模为 9 亿美元，其中基于扩增的检测约占总市场的 1/3。根据对潜在客户和诊断专业人员的调查，我们系统的市场潜力超过 1 亿美元。成功实现发展里程碑可以进一步推动这一潜力。