Allosteric DNAzyme sensors for practical detection of mycotoxins

用于实际检测霉菌毒素的变构 DNAzyme 传感器

• 批准号: 7106678
• 负责人: Scott K Silverman
• 金额: $ 10万
• 依托单位: DZYMETECH, INC.
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-02-01 至 2008-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7106678
• 关键词: Aflatoxin B1; Aflatoxins; Agriculture; Agrochemicals; Animals; Antibodies; Binding; Biological Assay; Biosensor; Businesses; Catalytic DNA; Cereals; Class; Cleaved cell; Condition; Containment; DNA; Dependence; Detection; Development; Domestic Animals; Economics; Engineering; Enzymes; Figs - dietary; Food Contamination; Food Supply; Fumonisin B1; Fumonisins; Goals; Gold; Growth; Health; Human; Illinois; In Vitro; Individual; Ions; Laboratories; Maize; Metals; Methods; Molds; Mycotoxicosis; Mycotoxins; National Security; Phase; Poison; Procedures; Process; Property; Protocols documentation; Public Health; Purpose; RNA; Relative (related person); Reporting; Research; Research Personnel; Safety; Sampling; Scientist; Security; Signal Transduction; Small Business Technology Transfer Research; Standards of Weights and Measures; System; Technology; Temperature; Testing; Toxin; Transportation; Universities; Wheat; Work; Zea mays; Zearalenone; base; deoxynivalenol; design; detector; experience; feeding; fluorophore; fungus; health economics; improved; innovation; interest; nanoparticle; practical application; programs; sensor; toe corn

DESCRIPTION (provided by applicant): Mycotoxins are toxic compounds produced by fungi that contaminate crops during growth, transportation, or storage. Contamination of food and feed by mycotoxigenic fungi is an important agricultural and national security concern. For example, thousands of people are sickened annually by mycotoxicoses related to crops contaminated with fungi, and the safety of the food supply is integral to our national health and security. All of these issues demand the development of improved toxin sensor technologies. Current mycotoxin detection methods (e.g., those based on antibodies) have practical shortcomings both in the development of the sensors and in their applications. In this Phase I STTR, we will develop innovative mycotoxin sensors based on allosteric DNAzymes, which are catalytic DNA molecules that are activated by binding of an analyte such as a mycotoxin. The long-term commercial objective of this project is to develop quantitative mycotoxin sensors for precise assays and also to develop semi-quantitative mycotoxin sensors for rapid field assays. This Phase I research will be performed with three specific aims: (1) Apply in vitro selection to identify allosteric DNAzymes that have high selectivity and sensitivity to the mycotoxin deoxynivalenol (DON) and other mycotoxins such as fumonisin B1, zearalenone (ZON), and aflatoxin B1; (2) Characterize and optimize the allosteric DNAzymes for toxin-sensing properties; and (3) Engineer fluorimetric or colorimetric mycotoxin sensors by integrating fluorophore or gold nanoparticle detection systems with the DNAzymes identified in Aims 1-2. In Phase II of the project, the allosteric DNAzyme sensors will be tested in the field to determine their absolute and relative efficacies as toxin detectors. This includes integration of the DNAzyme sensors with the standard storage and transport aeration system used in grain containment units, which will increase the assay sensitivities by concentrating analytical samples. Relevance to public health: Contamination of food and feed by mycotoxin-producing fungi is one of the most important health-related problems in the US and worldwide. Improved sensors for detecting mycotoxins and thereby revealing the presence of fungi will allow proper remedial actions to be taken.

描述（由申请方提供）：真菌毒素是由真菌产生的有毒化合物，在生长、运输或储存过程中污染作物。食品和饲料被真菌污染是一个重要的农业和国家安全问题。例如，每年有成千上万的人因与受真菌污染的作物有关的真菌中毒而患病，食品供应的安全是我们国家健康和安全的组成部分。所有这些问题都要求开发改进的毒素传感器技术。目前的真菌毒素检测方法（例如，基于抗体的那些）在传感器的开发和它们的应用中具有实际的缺点。在第一阶段STTR中，我们将开发基于变构DNA酶的创新真菌毒素传感器，变构DNA酶是通过结合分析物（如真菌毒素）激活的催化DNA分子。该项目的长期商业目标是开发用于精确测定的定量霉菌毒素传感器，并开发用于快速现场测定的半定量霉菌毒素传感器。本研究的第一阶段将进行三个具体目标：（1）应用体外筛选来鉴定对真菌毒素脱氧雪腐镰刀菌烯醇（DON）和其他真菌毒素如伏马菌素B1、玉米赤霉烯酮（ZON）和黄曲霉毒素B1具有高选择性和敏感性的变构DNA酶;（2）表征和优化变构DNA酶的毒素敏感特性;和（3）通过将荧光团或金纳米颗粒检测系统与目标1-2中鉴定的DNA酶整合，设计荧光或比色真菌毒素传感器。在该项目的第二阶段，变构DNA酶传感器将在现场进行测试，以确定其作为毒素检测器的绝对和相对功效。这包括将DNAzyme传感器与谷物容器中使用的标准储存和运输通风系统集成，这将通过浓缩分析样品来提高分析灵敏度。与公共卫生的相关性：产霉菌毒素真菌对食品和饲料的污染是美国和世界范围内最重要的健康相关问题之一。检测霉菌毒素的改进传感器，从而揭示真菌的存在，将允许采取适当的补救措施。