Innovative Biomonitoring Device for Rapid Diagnosis of Exposure to Nerve Agents

用于快速诊断神经毒剂暴露情况的创新生物监测装置

• 批准号: 7224666
• 负责人: Yuehe Lin
• 金额: $ 62.26万
• 依托单位: BATTELLE PACIFIC NORTHWEST LABORATORIES
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-30 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7224666
• 关键词: antibody; biomarker; blood; chelating agents; ions; metals; proteins; rapid diagnosis; saliva

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.

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，有必要开发新的和改进的对策来快速检测暴露。因此，本项目将开发一种便携式微量分析系统，用于快速评估有机磷（OP）神经毒剂的暴露，利用磷酸化胆碱酯酶（ChE）作为血液和唾液中的生物标志物（OP-ChE加合物）。我们的假设是，通过纳米颗粒连接抗体和金属螯合选择性捕获OP-ChE生物标志物，然后进行电化学检测，可以开发出一种高选择性、超灵敏的一次性生物传感器，用于检测亚临床暴露于神经毒剂。该项目直接响应RFA-NS-06-004，该RFA-NS-06-004涉及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响应能力的研究项目（U01）。该方法将包括针对OP-ChE生物标志物的免疫捕获电化学检测器的初步开发，以及随后的一次性免疫色谱电化学生物传感器（IEB）的开发。二异丙基氟化磷酸（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生物标记物将通过微流控IEB检测，使用类似ELISA检测的方法，不同之处在于，初级识别是由纳米颗粒标记的抗体进行的，二级识别是由氧化锆（ZrO2）与磷酸化片段螯合进行的，随后的检测是基于溶解纳米颗粒产生金属离子，并在金属离子积聚到电极上时检测金属离子。一种用于分析血液和/或唾液的实时生物监测IEB设备的开发代表了当前神经毒剂暴露生物监测策略的重大进步。微创生物基质的使用，加上现场和快速检测，代表了一种广泛应用于评估故意或意外暴露于化学神经毒剂的新方法。