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（由申请人提供）：分枝杆菌（MTB）对全球健康的影响令人震惊，每年约200万例死亡和约1000万例新的结核病病例（TB）。造成这一危机的主要贡献者是缺乏用于检测活性结核病的有效观察点（POC）诊断工具。这损害了临床医生的能力 为住院，隔离和治疗做出关键决定。结果，未诊断的患者发展为更严重的疾病，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TB的努力因多药耐药性结核（MDR-TB）的发病率的增加而变得复杂。尽管快速准确的药物敏感性测试（DST）对于控制活性结核病至关重要，但是在资源有限的设置中，DST廉价的POC诊断工具非常缺乏。该项目的目的是利用两部分脱氧核酶（DNAZYME）传感器的独特性能 - 高特异性和敏感性，低成本，在资源有限的设置中可行的实现 - 可显着提高MTB诊断功能。在AIM 1中，我们将设计和优化能够直接在痰液样品中直接对MTB RNA的超敏，物种特异性检测的DNAZYME传感器。在AIM 2中，我们将采用类似的技术来检测所选的单核苷酸多态性（SNP），这些突变已知可以赋予对临床相关的抗生素的抗性。最终，我们将寻求结合这两个工具 创建一个新颖的全面MTB诊断平台。这项技术有可能通过提供低成本，敏感且高度特定的POC分析来显着提高结核病诊断能力，并可以在资源有限的高累积环境中实施。

项目成果

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

• 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.