Rapid detection of methylglyoxal in blood samples

快速检测血液样本中的甲基乙二醛

• 批准号: 8455478
• 负责人: JOSEPH FRANCIS KREBS
• 金额: $ 16.17万
• 依托单位: BIOO SCIENTIFIC CORPORATION
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-01-22 至 2016-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8455478
• 关键词: Affect; American; Biological Assay; Biological Factors; Biological Markers; Blood; Blood Circulation; Blood Proteins; Blood Vessels; Blood specimen; Buffers; Cardiovascular Diseases; Cause of Death; Clinical; Complications of Diabetes Mellitus; Country; Coupled; Detection; Diabetes Mellitus; Diagnostic; Disease; Dyes; Enzymes; Equilibrium; Glucose; Goals; Health; Heart Diseases; High Pressure Liquid Chromatography; Housing; Hypertension; Incidence; Investigation; Lead; Life; Link; Measures; Mediating; Methods; Monitor; Mus; NADH; NADP; Neuropathy; Oxidoreductase; Pathogenesis; Phase; Plasma; Play; Preclinical Testing; Proteins; Pyruvaldehyde; Rattus; Reaction; Reader; Reagent; Recombinants; Research; Research Personnel; Rodent; Role; Sampling; Scientist; Serum; Signal Transduction; Testing; Time; Validation; absorption; analytical method; base; cardiovascular injury; combat; diabetic; glucose metabolism; microbial; novel; novel diagnostics; oxidation; pandemic disease; pre-clinical; prevent; public health relevance; rapid detection; sugar; tool

DESCRIPTION (provided by applicant): Diabetes is a serious, life-threatening disease caused by improper absorption and metabolism of glucose. Increased glucose levels can lead to an accumulation of toxic sugar degradation products, such as methylglyoxal (MG), in the bloodstream. MG is a reactive molecule which can modify and inactivate blood proteins which in turn leads to hypertension, neuropathy, and heart disease. Consequently, MG has been shown to be an excellent biomarker for a number of diabetes-related complications. Unfortunately, all current methods to directly measure MG levels in blood are very expensive, time-consuming and laborious. To fully realize the diagnostic potential of MG, rapid analytical methods are needed for the routine determination of MG in biomedical samples such as blood. Our proposed Phase I research will create a unique test to measure MG levels in blood samples such as plasma and serum. This unique test will permit researchers to conveniently study the accumulation of reactive MG in blood before damage to proteins and vasculature has occurred. Our novel assay will use a recombinant bacterial oxidoreductase enzyme (MG-R) capable of specifically and rapidly converting MG into a detectable colorimetric signal in a 96 well plate assay format. To boost sensitivity, we will link our enzymatic detection reaction to an NADP+/NADPH visible colorimetric amplification loop to increase assay sensitivity more than 100-fold. The assay will be optimized and validated using rat and mouse plasma and serum spiked with known amounts of MG. After validation, we will commercially launch a rapid test kit to directly detect MG in preclinical and other biomedical samples.

描述（由申请人提供）：糖尿病是一种严重的，威胁生命的疾病，原因是葡萄糖的吸收和代谢不当。葡萄糖水平升高会导致血液中有毒糖降解产物（例如甲基甘氨酸（MG））的积累。 MG是一种反应性分子，可以改变和失活血蛋白，进而导致高血压，神经病和心脏病。因此，MG已被证明是许多与糖尿病相关并发症的极好的生物标志物。不幸的是，所有直接测量血液中MG水平的方法都是非常昂贵，耗时且费力的。为了充分意识到MG的诊断潜力，在生物医学样本（如血液）中常规确定MG需要快速分析方法。我们提出的I期研究将创建一个独特的测试，以测量血浆和血清等血液样本中的MG水平。这项独特的测试将使研究人员可以方便地研究血液中反应型毫克的积累，然后才发生蛋白质和脉管系统。我们的新颖测定法将使用重组细菌氧化还原酶（MG-R），能够以96井板测定格式特异性地转化为可检测的比色信号。为了提高灵敏度，我们将将酶促检测反应与NADP+/NADPH可见的比色循环联系起来，以提高测定敏感性超过100倍。该测定法将通过大鼠和小鼠血浆和血清[已知量的Mg》进行优化和验证。验证后，我们将在商业上推出一个快速测试套件，以直接检测临床前和其他生物医学样品中的MG。