Rapid Susceptibility Testing of MDR M. tuberculosis

耐多药结核分枝杆菌的快速药敏试验

• 批准号: 6990908
• 负责人: MYRON SASSER
• 金额: $ 9.7万
• 依托单位: MIDI, INC.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2005-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6990908
• 关键词: Mycobacterium tuberculosis; bacteria characteristic; genetic strain; high performance liquid chromatography; hydroxy fatty acid; multidrug resistance; technology /technique development

DESCRIPTION (provided by applicant): This Phase I research will test the feasibility of using mycolic acid analysis as a very rapid test for drug susceptibility of Mycobacterium tuberculosis. The significance of TB is evident in that more than one-third of the world's population is infected by M. tuberculosis and multi-drug resistant TB is both a current serious health risk and potential bioterrorism weapon. Tuberculosis causes more death and suffering than any other infectious agent and is especially prevalent in HIV-infected populations. The current testing method for susceptibility of TB to treatment drugs averages nearly a week to obtain results and may be compromised by false results caused by bacterial contaminants. High performance liquid chromatography (HPLC) of mycolic acids enables rapid, inexpensive identification of mycobacteria as profiles are characteristic of the various species. When exposed to treatment drugs such as isoniazid, susceptible TB strains diminish in mycolic acid content within 20 minutes, as compared to the untreated controls, and resistant strains do not show this loss but gain in mycolic acid amount with time. Susceptible and resistant strains will be inoculated into control and inhibitor-containing vials of a commercial test system. The assay will be performed by PLC analysis of mycolic acid content over timed intervals (up to 72 h.). Comparative quantities of mycolic acids indicate growth/no growth and the characteristic pattern of mycolics assure that contaminants are not responsible for the result. As an alternative approach, intermediates/precursors such as hexacosanoic acid (C26:0) will be assayed using a similar research protocol and quantitation by HPLC analysis. PROPOSED COMMERCIAL APPLICATION: The major product will be the software and hardware necessary to perform the HPLC testing method. Internal standards and the HPLC Calibration mixture for the assay will also be commercial products. Phase II of the project will entail validating the methodology for multiple strains and additional antibiotics. Optimization for performing analyses directly from signal positive bottles and application to direct from sputum will be explored.

描述(申请人提供)：这项第一阶段的研究将测试使用霉酚酸分析作为一种非常快速的结核分枝杆菌药物敏感性测试的可行性。结核病的重要性是显而易见的，因为世界上三分之一以上的人口感染结核分枝杆菌，而耐多药结核病既是当前严重的健康风险，也是潜在的生物恐怖主义武器。结核病造成的死亡和痛苦比任何其他传染病都多，在感染艾滋病毒的人群中尤其普遍。目前的结核病对治疗药物敏感性的检测方法平均需要近一周的时间才能得出结果，而且可能会受到细菌污染造成的虚假结果的影响。分枝杆菌酸的高效液相色谱(HPLC)能够快速、廉价地鉴定分枝杆菌，因为不同物种的特征图谱是相同的。当暴露于异烟肼等治疗药物时，与未经处理的对照相比，敏感的结核病菌株在20分钟内霉菌酸含量减少，而耐药菌株没有表现出这种下降，但随着时间的推移真菌酸量增加。敏感和耐药菌株将被接种到商业测试系统的对照和含有抑制剂的小瓶中。该分析将通过PLC分析定时间隔(长达72小时)中的霉菌酸含量来进行。真菌酸的相对数量表明有生长/没有生长，真菌的特征模式确保了污染物不会对结果负责。作为一种替代方法，将使用类似的研究程序对中间体/前体进行分析，如十六烷酸(C26：0)，并通过高效液相分析进行定量。拟议的商业应用：主要产品将是执行高效液相测试方法所需的软件和硬件。该检测的内标和高效液相色谱校正混合物也将是商业化产品。该项目的第二阶段将需要对多种菌株和其他抗生素的方法学进行验证。将探索直接从信号阳性瓶进行分析的优化以及直接从痰中进行分析的应用。