Dipstick Assay for Detection of Acetaminophen Protein Adducts

描述（由申请人提供）：本STTR提案的最终目标是开发一种快速、敏感和特异性的诊断测试，可以识别人血清中对乙酰氨基酚（APAP）蛋白加合物。APAP毒性是当今美国和英国急性肝衰竭（ALF）最常见的原因。毒性是通过母体药物转化为反应性代谢物介导的，该代谢物以apap -蛋白加合物的形式与蛋白质共价结合。apap蛋白加合物作为毒性的结果出现在血清中，是一种很好的毒性生物标志物。急性APAP过量的诊断目前是通过接受毒性剂量后24小时内测定血清中APAP水平，或血清中肝转氨酶升高，并伴有毒性剂量史。这种方法有许多局限性，而且肝转氨酶的升高是非特异性的。对乙酰氨基酚毒性诊断有限责任公司将验证这一假设，即apap蛋白加合物可以使用基于抗体的试纸试验来确定。这将通过以下具体目标来实现：1)：开发一种用于测定apap蛋白加合物的快速原型试纸试验。2)：验证原型试纸法检测APAP过量患者血清中APAP-蛋白加合物的诊断敏感性和特异性。将试纸试验的结果与使用已建立的具有电化学检测的高效液相色谱法“平行”检测APAP蛋白加合物的结果进行比较。开发一种对APAP蛋白加合物敏感、快速的检测方法，将增强临床环境中APAP中毒的诊断，并导致早期识别有严重肝毒性风险的个体，从而降低发病率和死亡率。公共卫生相关性：在今天的美国，对乙酰氨基酚的过量使用是急性肝衰竭的一个主要原因。对乙酰氨基酚是许多用于治疗疼痛和发烧的非处方产品的主要成分。对乙酰氨基酚蛋白加合物是已知的对乙酰氨基酚引起的肝损伤的血液标志物。对乙酰氨基酚蛋白加合物的测量将有助于诊断对乙酰氨基酚相关的肝损伤，这些患者在过量服用24小时后出现在医疗中心。这是现有诊断测试不可靠的时间段。此外，对乙酰氨基酚蛋白加合物的测量可用于对乙酰氨基酚使用史不清楚或原因不明的急性肝衰竭患者。该项目将开发一种快速临床试验，用于测量对乙酰氨基酚蛋白加合物，可在当地医院实验室中使用，通过提高对对乙酰氨基酚的诊断和认识，将影响患者的整体医疗护理，并将减少对乙酰氨基酚毒性的未来死亡。

DESCRIPTION (provided by applicant): The ultimate goal of this STTR proposal is to develop a rapid, sensitive, and specific diagnostic test that can identify acetaminophen (APAP) protein adducts in human sera. APAP toxicity is the most common cause of acute liver failure (ALF) in the United States and Great Britain today. Toxicity is mediated by conversion of the parent drug to a reactive metabolite that covalently binds to protein as APAP-protein adducts. APAP-protein adducts appear in serum as a result of the toxicity and are an excellent biomarker of toxicity. The diagnosis of acute APAP overdose is currently made by determination of the APAP level in serum within 24 hours of receiving the toxic dose, or by elevation of hepatic transaminases in serum, accompanied by a history of toxic dosing. This approach has numerous limitations and the elevation of hepatic transaminases is nonspecific. Acetaminophen Toxicity Diagnostics, LLC will test the hypothesis is that APAP-protein adducts can be determined using an antibody-based dipstick assay. This will be accomplished through the following Specific Aims: 1): Develop a rapid prototype dipstick assay for determination of APAP-protein adducts. 2): Demonstrate the diagnostic sensitivity and specificity of the prototype dipstick assay for detection of APAP- protein adducts in the serum of APAP overdose victims. Results of the dipstick assay will be compared to "in parallel" examination of APAP protein adducts using an established high performance liquid chromatography assay with electrochemical detection. Development of a sensitive and rapid assay for APAP-protein adducts will enhance the diagnosis of APAP poisoning in clinical settings and lead to the earlier identification of individuals at risk for severe liver toxicity, thus reducing morbidity and mortality. PUBLIC HEALTH RELEVANCE: Overdoses of acetaminophen are a major cause of acute liver failure in the United States today. Acetaminophen is the major ingredient in many over- the - counter products sold for the treatment of pain and fever. Acetaminophen protein adducts are known blood markers of liver injury from acetaminophen. Measurement of acetaminophen protein adducts will help make the diagnosis of acetaminophen-related liver injury in patients that present to medical centers 24 hours after the overdose has occurred. This is a time period when the existing diagnostic test is not reliable. Also, measurement of acetaminophen protein adducts can be used in patients whose history of acetaminophen use is unclear or who have acute liver failure from unknown causes, This project will develop a rapid clinical test for measurement of acetaminophen protein adducts that can be used in local hospital laboratories and will affect the overall medical care of patients by improving the diagnosis and recognition of acetaminophen efforts and will reduce future deaths from acetaminophen toxicity.