Mass Spectrometry in Clinical Diagnosis of Nerve Agent Exposure

质谱法在神经毒剂暴露临床诊断中的应用

• 批准号: 7224047
• 负责人: OKSANA LOCKRIDGE
• 金额: $ 46.78万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-27 至 2009-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7224047
• 关键词: albumins; biomarker; blood; clinical research; diagnosis; human; mass spectrometry

DESCRIPTION (provided by applicant): The goal of this project is to develop a rapid and sensitive assay for nerve agent exposure in humans. Existing assays do not detect low dose exposure and cannot reassure the public that no exposure has occurred. Two proteins in blood, butyrylcholinesterase and albumin, will be tested as biomarkers of nerve agent exposure in our new assays. Butyrylcholinesterase is an established biomarker in assays that measure loss of enzyme activity. However, measurement of butyrylcholinesterase activity does not identify the inhibitor as a nerve agent, and does not reveal low dose exposure. Albumin is a new biomarker whose utility as a biomarker has not previously been tested in humans, though it shows promise in mice. Our proposed new assays use MALDI-TOF and tandem mass spectrometry to detect peptides covalently attached to nerve agent. The Selective Reaction Monitoring method is expected to detect as little as 100 femtomoles of covalently bound nerve agent in human plasma. The assays will be developed with human plasma treated with soman and sarin in vitro. The utility of the assays will be evaluated by testing plasma from human subjects exposed to nerve agent simulants. The specific aims are 1) Develop a method to rapidly diagnose exposure to nerve agents, by treating human blood with soman and sarin, and analyzing samples in the mass spectrometer. The method will look for peptides of albumin-nerve agent adducts and of butyrylcholinesterase-nerve agent adducts. 2) Develop a method to rapidly diagnose exposure to nerve agent simulants, by testing blood from humans accidentally exposed to organophosphorus pesticides, or from attempted suicide cases who ingested organophosphorus pesticides. Mass spectrometry will be used to identify exposure. This specific aim differs from aim #1 in that humans will have been exposed to nerve agent simulants; nerve agent simulants must be studied because humans exposed to nerve agents are not available. The new diagnostic methods have the potential for saving lives in an emergency because correct, rapid diagnosis indicates the appropriate medical treatment.

描述（由申请人提供）：该项目的目的是为人类的神经毒剂暴露开发快速敏感的测定法。现有测定未检测到低剂量的暴露，也不能向公众保证没有发生任何暴露。血液中的两种蛋白质，丁酰胆碱酯酶和白蛋白，将在我们的新测定中被视为神经毒剂暴露的生物标志物。在测量酶活性丧失的测定中，丁乙酸酯酶是一种已建立的生物标志物。但是，测量丁酰胆碱酯酶活性并不能识别抑制剂为神经剂，也没有发现低剂量的暴露。白蛋白是一种新的生物标志物，其作为生物标志物的实用性以前从未在人类中进行过测试，尽管它在老鼠中表现出了希望。我们提出的新测定法使用MALDI-TOF和串联质谱法检测与神经剂共同附着的肽。预计选择性反应监测方法将检测到人血浆中共价结合神经剂的100个替代性。这些测定将通过在体外用SOMAN和SARIN处理的人类血浆进行开发。测定的效用将通过测试暴露于神经剂模拟物的人类受试者的血浆来评估。具体目的是1）开发一种方法，通过用索曼和沙林治疗人体血液，并在质谱仪中分析样品，以快速诊断暴露于神经药物。该方法将寻找白蛋白障碍物加合物和丁酰胆碱酯酶 - 替代代理加合物的肽。 2）开发一种方法来快速诊断暴露于神经毒剂模拟物，通过意外暴露于有机磷农药的人或摄入有机磷农药的自杀案例的人类的血液中。质谱法将用于识别暴露。这个特定的目标与目标＃1不同，因为人类将暴露于神经剂模拟物中。必须研究神经毒剂模拟剂，因为没有暴露于神经剂的人。新的诊断方法有可能在紧急情况下挽救生命，因为正确，快速诊断表明适当的医疗。