DNA淀粉样蛋白传感器的分子标记与血浆和血糖因子,用于阿尔茨海默氏症的光子检测

R. Cuero, D. Agudelo, L. Sánchez, J. Londoño
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引用次数: 0

摘要

这项研究是基于细胞、分子和原子水平的综合方法进行的。因此,我们的目标是开发两种DNA传感器,利用合成生物学和分子生物学检测血液中与阿尔茨海默病相关的β-淀粉样蛋白。利用基因序列在大肠杆菌或酿酒酵母中构建DNA传感器,并在与人血浆混合时使用荧光检测器检测其荧光表达单位(FSU)。利用光子拉曼技术对该淀粉样蛋白传感器进行了验证,并利用SDS-PAGE凝胶电泳对纯化蛋白进行了分析,并利用不同尺寸的Macrosep Advance离心装置对该产品进行了验证。通过标记荧光染料偶联的荧光靶分子,增强了检测强度。荧光结果与临床和MRI结果相当。DNA传感器能够检测不同类型患者的β-淀粉样蛋白。结果还与患者的血糖水平相关。金属对淀粉样蛋白表达的影响也通过ELISA和Western Blot检测得到证实。由于荧光强度与β-淀粉样蛋白水平高度相关,这些结果被用于根据阿尔茨海默氏症的严重程度对患者进行分类(即组1:阿尔茨海默氏症诊断;Group2: Pre-Alzheimer的;组3:正常)、临床医学症状(如记忆丧失、认知障碍,如精神定向障碍和/或精神混乱)和MRI结果。统计分析表明,根据所选择的三个参数,可以很好地进行分组。第1组荧光总平均值最高,第2组次之,第3组荧光总平均值最低。性别似乎是疾病发病时的一个相关因素。这些结果表明使用这种方法早期检测阿尔茨海默病的优势以及金属对触发β-淀粉样蛋白表达的重要性。*通讯:Cuero R, PhD, 15310 Misty Dawn, Trl Cypress, TX, USA, Tel:(832) 477-5510;电子邮件:olimpa@aol.com
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Molecular Labeling of DNA Amyloid Sensor with Plasma, and the Glycemic Factor, for Photonic Detection of Alzheimer's in Blood
This investigation was carried out based on the integrative approach at the cellular, molecular and atomic level. Therefore, our aim was to develop two DNA sensors that detect Alzheimer’s disease related β-amyloid protein in blood using synthetic and molecular biology. The DNA sensors were constructed in E. Coli or Saccharomyces cerevisiae using genetic sequences and they were tested in terms of fluorescence expression units (FSU) when mixed with human blood plasma using a fluorescence detector. This amyloid sensor was also confirmed using photonic Raman technology, and corroborated by analyzing the purified protein using SDS-PAGE gel electrophoresis, which product was confirmed by using different sizes of Macrosep Advance Centrifugal devices. The intensity of the detection was enhanced by labeling the fluorescence targeted molecules in samples which were conjugated with fluorescent dyes. Fluorescence results were comparable to clinical and MRI results. The DNA sensor was able to detect β-amyloid protein in different type of patients. Results were also correlated with the glycemic levels of the patients. Influence of metals on expression of amyloid protein was also demonstrated through ELISA and Western Blot assays. Due to the highly correlation between fluorescence intensity and levels of β-amyloid, these results were used to classify patients according to the severity of Alzheimer’s (i.e. Group1: Alzheimer’s Diagnosed; Group2: Pre-Alzheimer’s; and Group3: Normal) in addition to clinical medical symptoms (e.g., memory loss, cognitive impairment such as mental disorientation and/or mental confusion) and MRI results. Statistical analysis showed that the groups were well categorized based on the three selected parameters. Group 1 showed the highest fluorescence total mean followed by Group 2, as compared to Group 3, which exhibited the lowest total mean. Gender seemed to be an associate factor at time of the disease onset. These results suggest the advantage of using this method for early detection of Alzheimer’s disease as well as the importance of metals for triggering the expression of β-amyloid protein. *Correspondence to: Cuero R, PhD, 15310 Misty Dawn, Trl Cypress, TX, USA, Tel: (832) 477-5510; E-mail: olimpa@aol.com
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