Innovative Biomonitoring Device for Rapid Diagnosis of Exposure to Nerve Agents

描述(由申请人提供)：随着涉及化学武器，包括神经毒剂的恐怖袭击的风险增加，有必要制定新的和改进的对策，以快速检测接触情况。因此，该项目将开发一种便携式微量分析系统，利用血液和唾液中的磷酸化胆碱酯酶(OP-CHE加合物)作为生物标志物，快速评估暴露于有机磷(OP)神经毒剂的情况。我们的假设是，通过使用纳米颗粒连接的抗体和金属螯合选择性地捕获OP-CHE生物标记物，然后进行电化学检测，可以开发出一种高选择性、超灵敏和一次性的生物传感器来检测亚临床暴露于神经毒剂。该项目直接响应RFA-NS-06-004关于对抗研究项目(U01)的要求，以开发新的诊断技术和工具，以提高我们在紧急情况下的医疗反应能力。该方法将包括初步开发用于OP-CHE生物标记物的免疫捕获电化学检测器，以及随后开发一次性免疫层析电化学生物传感器(IEB)。二异丙基磷酸盐(DFP)和沙林将作为化学神经毒剂在体外和体内暴露，在大鼠和人类的血浆、红细胞和唾液中产生OP-CHE蛋白生物标志物。OP-CHE生物标志物将通过微流控IEB使用类似于ELISA分析的方法进行检测，不同之处在于一次识别是通过纳米颗粒标记的抗体执行的，二次识别是通过氧化锆(ZrO2)与磷酸化部分的螯合来执行的，然后是基于通过溶解纳米颗粒产生金属离子并在积累到电极时检测金属离子的检测。用于血液和/或唾液分析的实时生物监测IEB设备的开发代表着相对于当前神经毒剂暴露的生物监测策略的重大进步。利用微创生物标记物，结合现场和快速检测，为评估故意或意外接触化学神经毒剂提供了一种新的方法，具有广泛的应用前景。

DESCRIPTION (provided by applicant): With an increased risk of terrorist attacks involving chemical weapons, including nerve agents, there is a need to develop new and improved countermeasures for the rapid detection of exposure. Hence, this project will develop a portable micro-analytical system for the rapid evaluation of exposure to organophosphorus (OP) nerve agents utilizing phosphorylated cholinesterase (ChE) as a biomarker (OP-ChE adducts) in blood and saliva. Our hypothesis is that a highly selective, ultra sensitive, and disposable biosensor can be developed to detect sub-clinical exposure to nerve agents by using nanoparticle linked antibodies and metal chelation to selectively capture OP-ChE biomarkers followed by electrochemical detection. This project is directly responsive to RFA-NS-06-004 concerning CounterACT research projects (U01) for the development of new diagnostic techniques and tools to enhance our medical response capabilities during an emergency. The approach will involve initial development of an immuno-capture electrochemical detector for OP-ChE biomarkers and the subsequent development of a disposable immunochromatographic electrochemical biosensor (IEB). Diisopropylphosphorofluoridate (DFP) and Sarin will be used as the chemical nerve agents in in vitro and in vivo exposures to generate OP-ChE protein biomarkers in the plasma, RBC, and saliva of rats and humans. OP-ChE biomarkers will be detected with the microfluidic IEB using an approach similar to the ELISA assay, except that primary recognition is performed by nanoparticle-tagged antibodies and secondary recognition is performed by zirconium oxide (ZrO2) chelation with the phosphorylated moiety, followed by detection that is based on generating metal ions by dissolving the nanoparticle and detecting the metal ions upon accumulation to an electrode. The development of a real-time biomonitoring IEB device for analysis of blood and/or saliva represents a significant advancement over current biomonitoring strategies for nerve agent exposure. The utilization of minimally invasive biomatrices, coupled with on-site and rapid detection represents a novel approach with broad application for evaluating deliberate or accidental exposures to chemical nerve agents.