Deoxyribozyme sensor-based diagnostics for Mycobacterium tuberculosis

基于脱氧核酶传感器的结核分枝杆菌诊断

• 批准号: 8574670
• 负责人: Dmitry Mikhaylovich Kolpashchikov
• 金额: $ 43万
• 依托单位: UNIVERSITY OF CENTRAL FLORIDA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8574670
• 关键词: Acid Fast Bacillae Staining Method; Address; Antibiotic Resistance; Antibiotics; Bacillus (bacterium); Bacteria; Base Sequence; Binding; Biological Assay; Catalytic DNA; Cessation of life; Child; Cleaved cell; Clinical; Detection; Development; Diagnostic; Disease; Disease Resistance; Drug resistance; Failure; Fatal Outcome; Gene Targeting; Goals; Gold; HIV Seropositivity; Hospitalization; Human; Incidence; Infection; Label; Left; Microarray Analysis; Microscopy; Multi-Drug Resistance; Multidrug-Resistant Tuberculosis; Mutation; Mycobacterium tuberculosis; Nucleic acid sequencing; Patients; Pharmaceutical Preparations; Point Mutation; Predisposition; Process; Property; RNA; Regimen; Reporter; Resistance; Resources; Ribosomal RNA; Rifampin; Sampling; Sensitivity and Specificity; Signal Transduction; Single Nucleotide Polymorphism; Solutions; Specificity; Sputum; Technology; Testing; Time; Translational Research; Tuberculosis; Vaccines; Validation; active control; base; clinically relevant; cost; design; disorder control; fluorophore; global health; graduate student; improved; isoniazid; killings; new technology; novel; novel diagnostics; point of care; point-of-care diagnostics; public health relevance; sensor; tool; undergraduate student

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.

描述(申请人提供)：结核分枝杆菌(Mtb)对全球健康的影响令人震惊，每年约有200万人死亡，约1000万新结核病(TB)病例。这场危机的一个主要原因是缺乏有效的护理点(POC)诊断工具来检测活动性结核病。这损害了临床医生的能力 做出关于住院、隔离和治疗的关键决定。因此，未确诊的患者会进展为更严重的疾病，并继续传播感染。更糟糕的是，目前涉及6-9个月多药方案的结核病治疗努力因耐多药结核病(MDR-TB)发病率的增加而变得复杂。尽管快速和准确的药敏试验(DST)对于控制活动性结核病至关重要，但在资源有限的情况下，用于DST的廉价POC诊断工具严重缺乏。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-高特异性和敏感度、低成本、在资源有限的情况下可行的实施-来显著提高结核分枝杆菌的诊断能力。在目标1中，我们将设计和优化能够在痰样本中直接检测Mtb RNA的超灵敏、特定物种的DNAzyme传感器。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(SNPs)，这些突变已知会导致对临床相关抗生素的耐药性。最终，我们将寻求将这两个工具结合起来 打造新型综合结核诊断平台。这项技术具有显著提高结核病诊断能力的潜力，它提供了一种可以在资源有限、高发环境中实施的低成本、高灵敏度和高特异性的POC检测方法。

项目成果

DNA Computing Systems Activated by Electrochemically-triggered DNA Release from a Polymer-brush-modified Electrode Array.

DNA计算系统是由电化学触发的DNA从聚合物刷子修饰的电极阵列释放而激活的。

• DOI: 10.1002/elan.201600389
• 发表时间: 2017-03
• 期刊: Electroanalysis
• 影响因子: 3
• 作者: Gamella M;Zakharchenko A;Guz N;Masi M;Minko S;Kolpashchikov DM;Iken H;Poghossian A;Schöning MJ;Katz E
• 通讯作者: Katz E

Nonequilibrium Hybridization Enables Discrimination of a Point Mutation within 5-40 °C.

• DOI: 10.1021/jacs.6b05628
• 发表时间: 2016-10-19
• 期刊: Journal of the American Chemical Society
• 影响因子: 15
• 作者: Stancescu M;Fedotova TA;Hooyberghs J;Balaeff A;Kolpashchikov DM
• 通讯作者: Kolpashchikov DM

Split Spinach Aptamer for Highly Selective Recognition of DNA and RNA at Ambient Temperatures.

• DOI: 10.1002/cbic.201600323
• 发表时间: 2016-09-02
• 期刊: Chembiochem : a European journal of chemical biology
• 作者: Kikuchi N;Kolpashchikov DM
• 通讯作者: Kolpashchikov DM

Divide and control: split design of multi-input DNA logic gates.

• DOI: 10.1039/c4cc08241a
• 发表时间: 2015-01-18
• 期刊: Chemical communications (Cambridge, England)
• 作者: Gerasimova YV;Kolpashchikov DM
• 通讯作者: Kolpashchikov DM

DNA Antenna Tile-Associated Deoxyribozyme Sensor with Improved Sensitivity.

• DOI: 10.1002/cbic.201600438
• 发表时间: 2016-11-03
• 期刊: CHEMBIOCHEM
• 影响因子: 3.2
• 作者: Cox, Amanda J.;Bengtson, Hillary N.;Gerasimova, Yulia V.;Rohde, Kyle H.;Kolpashchikov, Dmitry M.
• 通讯作者: Kolpashchikov, Dmitry M.