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（由申请人提供）：霉菌毒素是真菌产生的有毒化合物，在生长，运输或储存过程中污染了农作物。霉菌真菌对食物和饲料的污染是重要的农业和国家安全问题。例如，每年与被真菌污染的农作物有关的霉菌毒素每年都会使成千上万的人患病，而食物供应的安全是我们国家健康和安全的重要组成部分。所有这些问题都需要开发改进的毒素传感器技术。当前的霉菌毒素检测方法（例如，基于抗体的方法）在传感器及其应用中都有实际缺陷。在此阶段I STTR中，我们将基于变构含量的脱氧核糖质开发创新的霉菌毒素传感器，这些传感器是催化DNA分子，通过结合分析物（例如霉菌毒素）激活。该项目的长期商业目标是开发定量的霉菌毒素传感器以进行精确测定，并开发半定量霉菌毒素传感器以进行快速现场测定。这项I阶段研究将以三个特定的目的进行：（1）应用体外选择来鉴定对霉菌毒素脱氧烯醇（DON）和其他霉菌毒素（例如富莫尼酶B1）的选择性和敏感性的变构型dnazymes，例如富素蛋白B1，searalenone（Zon）（Zon）和Aflatoxin b1; （2）表征和优化毒素感应特性的变构虫； （3）通过将荧光团或金纳米颗粒检测系统与AIMS 1-2鉴定的Dnazymes相结合，通过将荧光团或金纳米颗粒检测系统整合到AIMS 1-2中，通过将荧光或比色霉菌毒素传感器进行了工程。在该项目的II阶段中，将在现场测试变构DNAZYME传感器，以确定其绝对和相对效率为毒素检测器。这包括将DNAZYME传感器与用于谷物遏制单元中的标准存储和传输曝气系统的整合，这将通过集中分析样品来提高测定敏感性。与公共卫生的相关性：产生霉菌毒素的真菌对食物和饲料的污染是美国和全球与健康相关的最重要问题之一。改进的传感器用于检测霉菌毒素，从而揭示了真菌的存在将允许采取适当的补救作用。