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通过分子印迹纳米粒子使用多种单体方法对麻风分枝杆菌进行表位印迹。

Epitope imprinting of Mycobacterium leprae bacteria via molecularly imprinted nanoparticles using multiple monomers approach.

机构信息

Department of Chemistry, MMV, Banaras Hindu University, Varanasi, 221005, India.

Department of Applied Science, Indian Institute of Information Technology, Allahabad, India.

出版信息

Biosens Bioelectron. 2019 Dec 1;145:111698. doi: 10.1016/j.bios.2019.111698. Epub 2019 Sep 12.

Abstract

Mycobacterium leprae causes endemic disease leprosy which becomes chronic if not treated timely. To expedite this 'timely diagnosis', and that also at an early stage, here an attempt is made to fabricate an epitope-imprinted sensor. A molecularly imprinted polymer nanoparticles modified electrochemical quartz crystal microbalance sensor was developed for sensing of Mycobacterium leprae bacteria through its epitope sequence. Multiple monomers, 3-sulphopropyl methacrylate potassium salt, benzyl methacrylate and 4-aminothiophenol were utilized to imprint this bacterial epitope. Imprinted nanoparticles were electropolymerized on gold coated quartz electrode. The sensor was able to show specific binding towards the blood samples of infected patients, even in the presence of 'matrix' and other plasma proteins such as albumin and globulin. Even other peptide sequences, similar to epitope sequences only with two amino acid mismatches were also unable to show any binding. Sensor withstood analytical tests viz. selectivity, specificity, matrix effect, detection limit (0.161 nM), quantification limit (and 0.536 nM), reproducibility (RSD 2.01%). Hence a diagnostic tool for bacterium causing leprosy is successfully fabricated in a facile manner which will broaden the clinical access and efficient population screening can be made feasible.

摘要

麻风分枝杆菌引起地方性疾病麻风病,如果不及时治疗,麻风病就会变成慢性疾病。为了加快这一“及时诊断”,并尽早进行诊断,这里尝试制造一种表位印迹传感器。通过麻风分枝杆菌的表位序列,开发了一种分子印迹聚合物纳米粒子修饰的电化学石英晶体微天平传感器,用于检测麻风分枝杆菌。利用多种单体,3-磺丙基甲基丙烯酸钾盐、苄基甲基丙烯酸酯和 4-氨基硫酚来印迹这种细菌的表位。印迹纳米粒子在金涂覆的石英电极上电聚合。该传感器能够特异性地结合感染患者的血液样本,即使在存在“基质”和其他血浆蛋白(如白蛋白和球蛋白)的情况下也是如此。即使是与表位序列只有两个氨基酸不匹配的其他肽序列,也无法显示任何结合。传感器经受住了分析测试,如选择性、特异性、基质效应、检测限(0.161nM)、定量限(和 0.536nM)、重现性(RSD 2.01%)。因此,以一种简单的方式成功地制造出了一种用于引起麻风病的细菌的诊断工具,这将扩大临床应用,并使有效的人群筛查成为可能。

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