Automatic Multi-Analyte In-Situ Bioassay for Monitoring Exposure to Toxic Metals

描述（由申请人提供） 一个现场便携式仪器，可以确定特定的有毒金属的血液水平是至关重要的建立有毒接触和基因表达之间的联系，因为它有助于识别基因的表达被修改的曝光。这些数据对实验室和流行病学研究都至关重要。标准的分析方法要求将样品送到场外实验室，这增加了成本和误差，并在确定接触源方面造成延误。拟议工作的总体目标将是开发一种小型、易于使用的手持式仪器，与一次性硅微芯片一起使用，以提供快速、现场同时检测血液中的铅、镉、铜和锌。微取样和分析芯片集成了一个尺寸与人的头发相当的微针，用于无痛抽血，带有一个含有电化学检测结构的微比色皿，可以使用MEMS（微机电系统）技术以成本效益高的方式制造。该分析将采用电化学溶出分析;这是一种经过验证的准确和精确检测痕量有毒重金属的技术。该测定允许检测分析物而不需要样品脱氧，并且使用内标物以避免需要外部校准。因此，它非常适合现场使用，并且可以由未经培训的人员使用。此外，样品体积的自计量将提供血液中四种金属水平的准确测定。第一阶段将通过以下方式证明其可行性：开发检测方法并通过体外测试进行验证，设计微芯片并制造关键部件，以及开发测量和控制仪器。在第二阶段，将制造一个完整的原型系统，并通过体外和动物试验进行评估。

DESCRIPTION (provided by applicant) A field-portable instrument that can determine the blood level of specific toxic metals is vital in establishing the link between toxic exposure and genetic expression in that it aides in identification of genes whose expression is modified by exposure. Such data is critical for both laboratory and epidemiological studies. Standard assay methods require samples be sent to off-site laboratories, increasing costs and errors, and imposing a delay in identifying exposure sources. The overall goal of the proposed work will be to develop a small, easy-to-use, handheld instrument used with disposable silicon microchips to provide rapid, in field simultaneous detection of lead, cadmium, copper and zinc in blood. The microsampling and assay chip, which integrates a microneedle comparable in size to a human hair for painless blood drawing with a microcuvette containing an electrochemical detection structure, can be cost-effectively fabricated using MEMS (microelectromechanical systems) technology. The assay will employ electrochemical-stripping analysis; a proven technique for accurate and precise detection of trace amounts of toxic heavy metals. The assay permits detection of the analytes without requiring sample de-oxygenation, and uses an internal standard to avoid the need for external calibration. It is thus ideal for field use and can be used by untrained personnel. In addition, self-metering of sample volume will provide for accurate determination of levels of the four metals in blood. Phase I will demonstrate feasibility by: developing the assay and validating it with in vitro testing, designing the microchip and fabricating key components, and developing measurement and control instrumentation. In Phase II, a complete prototype system will be fabricated and evaluated via in vitro and animal testing.