Deoxyribozyme sensor-based diagnostics for Mycobacterium tuberculosis

描述（申请人提供）：结核分枝杆菌（Mtb）对全球健康的影响是惊人的，每年约有200万人死亡，约有1000万新发结核病（TB）病例。造成这一危机的一个主要因素是缺乏用于检测活动性结核病的有效床旁（POC）诊断工具。这损害了临床医生的能力 做出有关住院、隔离和治疗的重要决定。因此，未确诊的患者进展为更严重的疾病，并继续传播感染。更糟糕的是，治疗结核病的努力，目前涉及6-9个月的多药方案，由于耐多药结核病（MDR-TB）发病率的增加而变得复杂。虽然快速准确的药物敏感性测试（DST）是控制活动性结核病的关键，但在资源有限的环境中，用于DST的廉价POC诊断工具非常缺乏。该项目的目标是利用双组分脱氧核酶（DNAzyme）传感器的独特特性-高特异性和灵敏度，低成本，在资源有限的环境中可行的实施-以显着提高结核病诊断能力。在目标1中，我们将设计和优化DNAzyme传感器，能够直接在痰液样本中超灵敏，种属特异性检测Mtb RNA。在目标2中，我们将应用类似的技术来检测选定的单核苷酸多态性（SNP），即已知会对临床相关抗生素产生耐药性的突变。最终，我们将寻求将这两种工具联合收割机结合起来 创建一个新型的结核病综合诊断平台。该技术具有通过提供低成本、灵敏度和高度特异性的POC测定来显著提高TB诊断能力的潜力，所述POC测定可以在资源有限的高发病率环境中实施。

DESCRIPTION (provided by applicant): The impact of Mycobacterium tuberculosis (Mtb) on global health is staggering, with ~2 million deaths and ~10 million new cases of tuberculosis (TB) each year. A major contributor to this crisis is the lack of effective point- of-care (POC) diagnostic tools for the detection of active TB disease. This compromises the ability of clinicians to make critical decisions regarding hospitalization, isolation, and treatment. As a result, undiagnosed patients progress to more severe disease and continue to transmit the infection. To make matters worse, efforts to treat TB, which currently involves a 6-9 month multi-drug regimen, are complicated by the increasing incidence of multidrug resistant TB (MDR-TB). Although rapid and accurate drug susceptibility testing (DST) is crucial for the control of active TB, an inexpensive POC diagnostic tool for DST in resource-limited settings is sorely lacking. The goal of this project is to exploit the unique properties of two-component deoxyribozyme (DNAzyme) sensors - high specificity and sensitivity, low cost, feasible implementation in resource-limited settings - to dramatically improve Mtb diagnostic capabilities. In Aim 1, we will design and optimize DNAzyme sensors capable of ultrasensitive, species-specific detection of Mtb RNA directly in sputum samples. In Aim 2, we will apply a similar technology to detect selected single nucleotide polymorphisms (SNPs), mutations that are known to confer resistance to clinically relevant antibiotics. Ultimately, we would seek to combine these two tools to create a novel comprehensive Mtb diagnostic platform. This technology has the potential to significantly improve TB diagnostic capabilities by providing a low-cost, sensitive, and highly specific POC assay that could be implemented in resource limited, high-incidence settings.