Helmholtz Centre for Environmental Research (UFZ), Permoserstraße 15, Leipzig 04318, Germany.
Sensors (Basel). 2014 Feb 21;14(2):3737-55. doi: 10.3390/s140203737.
Aptamers are short, single stranded DNA or RNA oligonucleotides that are able to bind specifically and with high affinity to their non-nucleic acid target molecules. This binding reaction enables their application as biorecognition elements in biosensors and assays. As antibiotic residues pose a problem contributing to the emergence of antibiotic-resistant pathogens and thereby reducing the effectiveness of the drug to fight human infections, we selected aptamers targeted against the aminoglycoside antibiotic kanamycin A with the aim of constructing a robust and functional assay that can be used for water analysis. With this work we show that aptamers that were derived from a Capture-SELEX procedure targeting against kanamycin A also display binding to related aminoglycoside antibiotics. The binding patterns differ among all tested aptamers so that there are highly substance specific aptamers and more group specific aptamers binding to a different variety of aminoglycoside antibiotics. Also the region of the aminoglycoside antibiotics responsible for aptamer binding can be estimated. Affinities of the different aptamers for their target substance, kanamycin A, are measured with different approaches and are in the micromolar range. Finally, the proof of principle of an assay for detection of kanamycin A in a real water sample is given.
适体是短的、单链 DNA 或 RNA 寡核苷酸，能够特异性地结合高亲和力到他们的非核酸靶分子。这种结合反应使它们能够作为生物识别元件在生物传感器和分析中应用。由于抗生素残留造成的问题有助于抗生素耐药病原体的出现，从而降低药物对抗人类感染的有效性，我们选择了针对氨基糖苷类抗生素卡那霉素 A 的适体，目的是构建一个强大的和功能的分析方法，可以用于水分析。通过这项工作，我们表明，从针对卡那霉素 A 的 Capture-SELEX 程序中衍生的适体也显示与相关的氨基糖苷类抗生素结合。所有测试的适体之间的结合模式不同，因此有高度物质特异性的适体和更多的群体特异性适体结合到不同种类的氨基糖苷类抗生素。也可以估计负责适体结合的氨基糖苷类抗生素的区域。不同适体对其靶物质，卡那霉素 A 的亲和力用不同的方法进行测量，在微摩尔范围内。最后，给出了在实际水样中检测卡那霉素 A 的分析检测的原理证明。
