Specific Diagnostic for Members of Genus Mycobacterium

分枝杆菌属成员的特异性诊断

• 批准号: 6933252
• 负责人: RICHARD Ashley HURT
• 金额: $ 10万
• 依托单位: ATOM SCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-06-01 至 2006-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6933252
• 关键词: Mycobacterium; Mycobacterium tuberculosis; bacterial DNA; clinical research; diagnostic tests; human tissue; microarray technology; nucleic acid sequence; respiratory disorder diagnosis; technology /technique development

DESCRIPTION (provided by applicant): A novel microarray-based system is proposed for diagnosis of tuberculosis and tuberculosis-like respiratory infections. This diagnostic system offers a parallel method to screen clinical samples for the presence of DNA sequences from Mycobacterium tuberculosis, the causative agent of human tuberculosis, and closely related mycobacterial species, while simultaneously screening specimens for more distantly related mycobacteria and other common infectious agents known to cause tuberculosis-like symptoms. The system selectively amplifies targeted DNA sequences using repeated rounds of primer-directed extension. Unlike the PCR, this method generates short extension products whose lengths are limited by using one of the building blocks in a form that terminates the extension reaction when it is incorporated. Restricting the extension reaction to the production of short extension products allows screening for many pathogens with a single reaction. The technique is very selective because a signal is produced only when the extension reaction products hybridize to probes immobilized on the microarray. This requires that: 1), the primer binds with a complementary sequence on the targeted DNA, and 2) the resulting short extension sequence is complementary to the probe on the microarray used to detect the extension product. Both of these are very selective processes.

描述（由申请人提供）：提出了一种用于诊断结核病和结核样呼吸道感染的新型基于微阵列的系统。该诊断系统提供了一种平行的方法来筛选临床样品中是否存在来自结核分枝杆菌（人类结核病的病原体）和密切相关的分枝杆菌物种的DNA序列，同时筛选标本中是否存在更远的相关分枝杆菌和已知引起结核样症状的其他常见感染因子。该系统使用重复轮的引物定向延伸选择性地扩增靶向DNA序列。与PCR不同，该方法产生短的延伸产物，其长度通过使用一种结构单元来限制，该结构单元的形式是当其被掺入时终止延伸反应。将延伸反应限制于产生短延伸产物允许用单一反应筛选许多病原体。该技术的选择性很强，因为只有当延伸反应产物与固定在微阵列上的探针杂交时才产生信号。这就要求：1）引物与靶DNA上的互补序列结合，和2）所得短延伸序列与用于检测延伸产物的微阵列上的探针互补。这两个过程都是非常有选择性的。