Easy, fast, point of care nucleic acid based detection of pulmonary tuberculosis

简单、快速、即时的基于核酸的肺结核检测

• 批准号: 7108320
• 负责人: Heather Koshinsky
• 金额: $ 9.96万
• 依托单位: INVESTIGEN, INC.
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-06-01 至 2006-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7108320
• 关键词: fasting; nucleic acids; sputum; tuberculosis

DESCRIPTION (provided by applicant): The proposed research is to determine the feasibility of developing a novel, simple, rapid method of detecting the nucleic acid (NA) from Mycobacterium tuberculosis cell in sputum, for detection of pulmonary tuberculosis (TB). This method would be based on Investigen's novel and elegant "smartDNA" technology. This system utilizes a peptide nucleic acid (PNA) probe cocktail in conjunction with a dye and light activation to produce a rapid color change when probes hybridize to complementary NA sequences. This color change can be visualized by the naked eye or detected by an absorbance or fluorescence reader. The four aims of feasibility are 1) evaluate the use synthetic sputum and M. microti as a surrogate for human sputum and M. tuberculosis; 2) develop a sputum processing protocol that is compatible with smartDNA-based detection of TB; 3) determine specificity and sensitivity of the entire sputum processing- smartDNA TB detection system in the detection of M. tuberculosis NA and 4) validate that our simple, rapid lysis method releases NA from M. tuberculosis. With this validation of the feasibility of developing a complete smartDNA TB detection system, we would move to Phase II objectives and test the system with normal human sputum samples spiked with M. tuberculosis and with clinical samples. The intention is to create a commercial diagnostic assay to detect TB in patients. Such a diagnostic could be used in peripheral locations, away from a central lab. This is greatly needed and would help curb the spread of TB by offering more rapid treatment. The proposed research would use a surrogate system to examine the feasibility of developing a method for the rapid detection of M. tuberculosis out of sputum. Findings from this study would be very useful in developing a novel commercial test for patients with active TB.

描述（由申请人提供）：拟议的研究是为了确定开发一种新的、简单的、快速的方法来检测痰中结核分枝杆菌细胞的核酸（NA），用于检测肺结核（TB）的可行性。这种方法将基于Investigen公司新颖而优雅的“智能dna”技术。该系统利用肽核酸（PNA）探针鸡尾酒结合染料和光激活，当探针与互补的NA序列杂交时，产生快速的颜色变化。这种颜色变化可以用肉眼观察，也可以用吸光度仪或荧光仪检测。可行性研究的四个目的是：1)评价人工痰液和微分枝杆菌作为人类痰液和结核分枝杆菌的替代品；2)开发与基于智能dna的结核病检测兼容的痰液处理方案；3)确定整个痰液处理- smartDNA结核检测系统检测结核分枝杆菌NA的特异性和敏感性；4)验证我们的简单，快速裂解方法从结核分枝杆菌中释放NA。在验证了开发完整的smartDNA结核病检测系统的可行性后，我们将进入II期目标，并在含有结核分枝杆菌的正常人类痰样本和临床样本中测试该系统。其目的是创建一种商业诊断方法来检测患者的结核病。这样的诊断可以在远离中心实验室的外围位置使用。这是非常需要的，并且将通过提供更快速的治疗来帮助遏制结核病的传播。拟议的研究将使用替代系统来检查开发一种快速检测痰液中结核分枝杆菌的方法的可行性。这项研究的结果将对开发一种针对活动性结核病患者的新型商业测试非常有用。

项目成果

Accelerated photobleaching of a cyanine dye in the presence of a ternary target DNA, PNA probe, dye catalytic complex: a molecular diagnostic.

• DOI: 10.1021/ac702519k
• 发表时间: 2009-03-15
• 期刊: ANALYTICAL CHEMISTRY
• 影响因子: 7.4
• 作者: Wang, M.;Holmes-Davis, R.;Rafinski, Z.;Jedrzejewska, B.;Choi, K. Y.;Zwick, M.;Bupp, C.;Izmailov, A.;Paczkowski, J.;Warner, B.;Koshinsky, H.
• 通讯作者: Koshinsky, H.