Miniarray Detection of MTB Complex Species

描述(申请人提供)：结核病是全球最重要的单一病原体致死原因。诊断主要通过对染色的痰涂片和用于生物测试的培养物进行显微镜分析进行。虽然染色很快，但不确定，虽然培养是确定的，但速度很慢。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用于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分析的速度和灵敏度优势，但使用了一种名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，并在一次测试中提供了多个物种的区分。这种微型阵列平台的特点还有望通过使用专有的信号放大方法和试剂，在较新版本的分析中克服对PCR扩增的需求。因此，预计将有两条产品线：1.为发达国家提供高技术版本，以提供更强的诊断能力以及更快、更可靠的一线检测；2.为发展中国家提供低技术桌面分析版本，能够快速区分多种物种，但无需聚合酶链式反应设备、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，以区分主要的结核分枝杆菌复杂亚种。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特定物种诊断模式，反映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缺失。第二阶段将把这些产品推向市场，将它们推广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型阵列。 这项技术基于专利的、正在申请专利的试剂和程序，这些试剂和程序将基因组DNA样本转化为用通用连接子修饰的放大的目标拷贝。然后，这些特殊的连接子能够用一套单一的引物对这组靶标进行全局扩增，它们还通过结合被称为GeneTAGs的基于DNA的长报告基因来提供增加信号传递的手段。相关的微型阵列平台还提供了一条开发各种类似诊断生物芯片的途径，用于区分其他病原体。这种小型阵列系统将快速(&lt；2小时)、灵敏(相当于聚合酶链式反应)、廉价且易于执行。全球结核病检测市场价值9亿美元，其中以扩增为基础的检测约占总数的三分之一。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能会推动这一潜力的提高。

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.