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）：结核病是世界范围内单一病原体导致死亡的最重要原因。诊断主要是通过显微镜分析染色的痰涂片和生长培养物进行生物试验。染色快速，但不确定；培养明确，但缓慢。基于核酸扩增的专有检测快速可靠，但它们需要的设备和试剂只适合发达国家的先进实验室。提出了一种新的微型阵列检测系统，它结合了基于扩增的检测方法的速度和灵敏度优势，但采用了一种简化的微阵列检测平台，称为微型阵列，降低了成本和复杂性，并在一次测试中提供了多种物种的区分。该微型阵列平台的特点还承诺，通过采用专有的信号扩增方法和试剂，克服对PCR扩增的需要。因此，预计将有两条产品线：一种面向发达国家的高科技版本，以提供更强的诊断能力以及快速、更可靠的一线检测。这是一种面向发展中国家的低技术含量的台式分析版本，可以快速区分多种物种，但无需PCR设备、高分辨率微阵列扫描仪或荧光显微镜即可进行诊断。第一阶段将重点开发和测试两个用于区分结核分枝杆菌主要复杂亚种的原型微型阵列。虽然最初的微型阵列原型仍然基于PCR扩增，但它们将展示视觉上简单的、物种特异性的诊断模式，反映出一系列相关但离散的物种或亚种特征的关键基因片段的存在和缺失。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，并将开始为发展中国家开发类似的低技术微型阵列。