Development of a phage-based nucleic acid amplification diagnostic for the rapid

描述（申请人提供）：1993年4月，世界卫生组织（WHO）宣布结核病（TB）为全球卫生紧急情况，这是从未给予其他疾病的区别。20亿人感染结核分枝杆菌（Mtb），每年导致800万新的结核病病例，近300万人因此死亡。尽管开发了有效的化疗来治疗结核病，但这一公共卫生危机仍然存在，并受到艾滋病毒共同流行的推动。 结核病控制的一个重大障碍是抗药性疾病的出现。耐多药结核菌株（MDRTB）在全世界约5000万人中持续存在，每年导致近50万例MDRTB新病例。耐多药结核病比药物敏感性结核病更难治疗，治疗费用也更高，需要长达两年的药物组合治疗，包括比一线疗法效力更低、毒性更大的二线药物。在这场公共卫生危机中，一个更令人不安的事态发展是广泛耐药结核菌株的全球分布，这可能源于对耐多药结核病例的管理不善。这些XDR菌株对异烟肼和利福平以及关键的二线药物具有耐药性;因此，它们甚至比MDRTB更难治疗，并且通常对感染者来说是死刑。 我们建议开发一种新的快速诊断结核病的抗生素敏感性测试，结合了两种成熟技术的最佳特征。第一个是分枝杆菌噬菌体在药物治疗后仅特异性感染活的和有免疫能力的分枝杆菌的能力;检测来自噬菌体的翻译产物然后可以用作抗生素抗性的标记。第二是各种核酸扩增和检测技术的灵敏度，这些技术只能检测特定序列的几个拷贝。我们希望这两种技术的结合能够非常敏感和特异地确定结核病的抗生素敏感性。此外，我们相信，它显示出很大的希望，能够直接进行痰液标本。 每年有800万新的结核病病例，造成近300万人死亡。结核病控制的一个重大障碍是抗药性疾病的出现。在这项拨款申请中，我们建议开发一种新的方法来检测耐药结核病，这种方法比现有方法更快，更便宜，使耐药疾病患者能够立即用适当的药物进行治疗，以防止其进一步传播。

DESCRIPTION (provided by applicant): In April 1993, the World Health Organization (WHO) declared tuberculosis (TB) a global health emergency, a distinction never accorded another disease. Two billion people are infected with Mycobacterium tuberculosis (Mtb), leading to eight million new TB cases each year and nearly three million resultant deaths. Despite the development of effective chemotherapies to treat TB, this public health crisis persists, fueled by the burgeoning HIV co-epidemic. A considerable obstacle to TB control is the emergence of drug-resistant disease. Multidrug-resistant strains of TB (MDRTB) persist in approximately 50 million people worldwide and lead to nearly 500,000 new cases of MDRTB each year. MDRTB is much more difficult and expensive than drug-susceptible TB to treat, and requires up to two years of therapy with a combination of drugs, including second-line drugs that are less potent and more toxic than the first-line therapies. An even more disquieting development in this public health crisis is the global distribution of strains of extensively drug-resistant TB (XDRTB), which may originate from poor management of MDRTB cases. These XDR strains are resistant to isoniazid and rifampin, as well as to key second-line drugs; as a result, they are even more difficult to treat than MDRTB, and are often a death sentence for those infected. We propose to develop a novel rapid diagnostic for antibiotic susceptibility testing for TB that combines the best characteristics of two proven technologies. The first is the ability of mycobacteriophage to specifically infect only viable and translationally competent Mycobacteria following drug treatment; detection of a translated product from the phage can then serve as a marker for antibiotic resistance. The second is the exquisite sensitivity of various nucleic acid amplification and detection technologies, which have the capability to sense only a few copies of a particular sequence. Our hope is that the marriage of these two technologies will allow for the very sensitive and specific determination of the antibiotic susceptibility profile of TB. Further, we believe that it shows great promise for being able to be performed directly on sputum specimens. There are eight million new cases of tuberculosis (TB) each year and nearly three million resultant deaths. A considerable obstacle to TB control is the emergence of drug-resistant disease. In this grant application, we propose to develop a novel method for the detection of drug-resistant TB that would be significantly faster and less expensive than existing methods, allowing patients with drug-resistant disease to be immediately treated with appropriate drugs to prevent its further transmission.