Catalytic DNA Biosensor for Toxic Metal Ions

描述（由申请人提供）：将开发用于重金属离子如铅（II）、铀（VI）、汞（II）和镉（II）的具有成本效益的催化DNA生物传感器。将对它们进行现场、实时检测和定量测试，假阳性或假阴性结果的发生率最低。暴露于这些金属离子会对人类，特别是儿童造成严重的不良健康影响，例如对脑、肾的损害和癌症。目前的金属检测技术采用昂贵或复杂的仪器或简单的传感器，其中许多具有高的假阳性或假阴性结果率。因此，迫切需要可靠的便携式重金属传感器，用于政府检查员、环境监测员、家庭用户、农村或偏远地区的临床医生以及反恐战争中的第一反应者，其中现场、实时和具有成本效益的检测和量化是至关重要的，并且重金属检测的市场估计为> 6.5亿美元。该项目基于伊利诺伊大学PI小组的几项创新，利用了催化DNA生物学，催化分子信标生物技术和纳米颗粒纳米技术的最先进工具。将通过体外选择获得对重金属离子具有特异性的催化DNA分子，并通过将DNA与荧光团/猝灭剂对和金纳米颗粒分别结合，将其转化为荧光（用于定量测量）和比色（用于定性和半定量测量）传感器。在NIH STTR第一阶段项目（1 R41 ES 014125 -01）中已经确定了在DzymeTech将创新转化为商业成功的可行性，其中已经获得了具有高灵敏度的铅（II）和铀（VI）催化DNA传感器（例如，~ 11 ppt）和选择性（例如，> 100万倍）。该第二阶段应用解决了将传感器设计转化为市场传感器套件原型的关键问题。将确定客户要求，并开发原型组件和协议。它们包括：“获得Pb（II）、U（VI）、Cd（II）和Hg（II）的所有四种荧光和比色传感器。通过确定每个传感器的检测限和选择性来表征传感器。“为每种金属离子设计和表征原型传感器套件。包括传感器组件、储存条件和测试方案优化、高/低温稳定性测试以及多批次传感器的重现性测试。“测试现场样本以研究干扰和基质效应。将收集和测试真实的现场样本，包括土壤和水样本。使用真实的样本的探头性能将用于评价探头套件的商业化价值。 公共卫生相关性：将开发用于铅（II）、铀（VI）、镉（II）和汞（II）的高灵敏度和选择性催化DNA传感器，用于具有成本效益的现场和实时检测和定量。重金属离子广泛存在于环境中，对人类尤其是儿童的脑、肾和神经系统造成严重的健康问题。该项目开发的方便便携式传感器将使政府检查员、环境监测员、家庭用户、临床医生和急救人员能够就补救和监测策略做出明智的决定，从而改善人类健康和生活质量。

DESCRIPTION (provided by applicant): Cost-effective catalytic DNA biosensors for heavy metal ions such as lead(II), uranium(VI), mercury(II), and cadmium(II) will be developed. They will be tested for on-site, real-time detection and quantification with minimal rates of false positive or false negative results. Exposure to these metal ions can cause severe adverse health effects, such as damages to brain, kidneys, and cancers, to human beings, especially children. Current technologies for metal detection employ either expensive or sophisticated instruments or simple sensors, many of which have high rates of false positive or false negative results. Therefore there is an urgent need of reliable portable heavy metal sensors for government inspectors, environmental monitors, household users, clinicians in rural or remote areas, and first-responders in the war against terrorism, where on-site, real-time, and cost-effective detection and quantification are critical and the market for heavy metal detection is estimated to be >$650 million. The project is based on several innovations in the PI's group at the University of Illinois that takes advantage of state-of-the-art tools in catalytic DNA biology, catalytic molecular beacons biotechnology and nanoparticle nanotechnology. Catalytic DNA molecules specific for heavy metal ions will be obtained through in vitro selection, and converted to fluorescent (for quantitative measurement) and colorimetric (for qualitative and semi-quantitative measurement) sensors by combining the DNA with fluorophore/quencher pairs and gold nanoparticles, respectively. The feasibility of transforming the innovations into commercial success at the DzymeTech has been established in the NIH STTR Phase I project (1R41ES014125-01), in which catalytic DNA sensors for lead(II) and uranium(VI) have been obtained that have high sensitivity (e.g., ~ 11 ppt) and selectivity (e.g., > 1 million fold). This Phase II application addresses critical issues of transferring the sensor design into prototypes of sensor kits for markets. The customer requirements will be determined and prototype components and protocols will be developed. They include: "Obtain all four fluorescent and colorimetric sensors for Pb(II), U(VI), Cd(II), and Hg(II). Characterize the sensors by determining the detection limit and selectivity of each sensor. "Design and characterize prototype sensor kits for each metal ion. Including sensor components, storage condition and test protocol optimization, high/low temperature stability tests and reproducibility tests with sensors from multiple-batches. "Test field samples to study interference and matrix effects. Real field samples, including soil and water samples will be collected and tested. Sensor performance with real samples will be used to evaluate the commercialization values of the sensor kits. PUBLIC HEALTH RELEVANCE: Highly sensitive and selective catalytic DNA sensors for lead(II), uranium(VI), cadmium(II), and mercury(II) will be developed for cost-effective on-site and real-time detection and quantification. Heavy metal ions are widely present in the environment, causing significant health problems such as damages to brain, kidney, and neural system to human beings, especially children. Convenient portable sensors developed in this project will enable government inspectors, environmental monitors, household users, clinicians and first-responders make informed decision about remediation and monitoring strategy and thus improve human health and quality of life.