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使用炔基修饰的葡聚糖底物比色法检测叠氮胸苷

Colorimetric Detection of Azidothymidine Using an Alkyne-Modified Dextran Substrate.

作者信息

Pratt George W, Fan Andy, Klapperich Catherine M

机构信息

College of Engineering, Boston University, Boston, Massachusetts 02215, United States.

出版信息

ACS Biomater Sci Eng. 2015 May 11;1(5):314-319. doi: 10.1021/ab500130d. Epub 2015 Apr 23.

Abstract

Monitoring the adherence of patients taking highly active antiretroviral therapy (HAART) is a key step in treating an HIV infection, especially in resource-limited settings in the developing world. For most regimens, when patients are not at least 95% adherent to their drug schedule, there is a loss of effectiveness in treatment resulting in increases in health care costs, increases in the rate of transmission, and reduction of positive patient outcomes. Currently, subjective methods such as pill counting, electronic drug monitoring, and patient self-reporting are the only ways clinicians can track adherence and intervene in cases of noncompliance. We address this issue by developing a dipstick-based point-of-care azide-alkyne click chemistry assay with colorimetric read-out that directly tests for the presence of one common HAART drug in patient urine. An alkyne-modified dextran was synthesized and characterized by NMR and then used to colorimetrically report the presence of azidothymidine, an azide-containing HAART drug, in urine samples. The assay is specific to azide-containing molecules that are not naturally present in the urine and is sensitive to physiologically relevant urine concentrations as low as 750 uM. This point-of-care device is a strong alternative in resource-limited settings over current direct measurement techniques that are expensive and require trained users such as HPLC.

摘要

监测接受高效抗逆转录病毒疗法(HAART)的患者的依从性是治疗HIV感染的关键步骤,在发展中国家资源有限的环境中尤为如此。对于大多数治疗方案,当患者对药物治疗方案的依从性低于95%时,治疗效果会降低,导致医疗成本增加、传播率上升以及患者积极治疗结果减少。目前,诸如药丸计数、电子药物监测和患者自我报告等主观方法是临床医生跟踪依从性并对不依从情况进行干预的唯一途径。我们通过开发一种基于试纸条的即时检测叠氮化物 - 炔烃点击化学分析法来解决这个问题,该分析法采用比色读出,可直接检测患者尿液中一种常见的HAART药物的存在。合成了一种炔烃修饰的葡聚糖并用核磁共振进行了表征,然后用于比色报告尿液样本中含叠氮化物的HAART药物叠氮胸苷的存在。该分析方法对尿液中天然不存在的含叠氮化物分子具有特异性,并且对低至750 μM的生理相关尿液浓度敏感。在资源有限的环境中,这种即时检测设备是当前直接测量技术(如HPLC,既昂贵又需要训练有素的用户)的有力替代方案。

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