Department of Molecular Biology, Institute of Nanosensor and Biotechnology, Dankook University, Yongin 448-701, Republic of Korea.
Department of Chemistry and Education, Seoul National University, Seoul 151-742, Republic of Korea.
Biochem Biophys Res Commun. 2014 Jun 13;448(4):397-402. doi: 10.1016/j.bbrc.2014.04.103. Epub 2014 May 2.
Danofloxacin is a synthetic fluoroquinolone with broad spectrum antibacterial activity that is used for the treatment of respiratory diseases in animal husbandry. However, danofloxacin has many adverse reactions and is toxic to humans. Especially, it detrimentally affects muscle, central nerve system, peripheral nerve system, liver, and skin in those who ingest foods in which danofloxacin has accumulated. Prescreening and determination of the level of danofloxacin in foods or food products is necessary for human health. Aptamers are composing of oligonucleotides that specifically interact with target molecules. They are emerging as detection/diagnostic ligands. Here, we used the SELEX in vitro selection technology to identify specific and high-affinity RNA aptamers with 2'-fluoro-2'-deoxyribonucleotide modified pyrimidine nucleotides against danofloxacin. Selected RNA aptamers bound specifically to danofloxacin, but not to tetracycline. Truncation of RNA aptamer up to 36 mer did not comprise specificity and affinity. The truncated RNA aptamer specifically bound to target chemical, allowing the discrimination of danofloxacin from other fluoroquinolones. The isolated specific aptamer could be a potential agent used for the rapid and cost-effective detection and sensing of danofloxacin, replacing instrumental methods including the more expensive and time-consuming methods of high performance liquid chromatography and liquid chromatography/mass spectrometry.
丹氟沙星是一种具有广谱抗菌活性的合成氟喹诺酮类药物,用于治疗畜牧业中的呼吸道疾病。然而,丹氟沙星有许多不良反应,对人类有毒性。特别是,它会对摄入含有丹氟沙星的食物的人的肌肉、中枢神经系统、周围神经系统、肝脏和皮肤造成损害。为了人类健康,有必要对食品或食品中丹氟沙星的含量进行筛选和测定。适体是由特定与靶分子相互作用的寡核苷酸组成的。它们正成为检测/诊断配体。在这里,我们使用体外 SELEX 选择技术,鉴定出针对丹氟沙星的具有 2'-氟-2'-脱氧核糖核苷酸修饰嘧啶核苷酸的特异性和高亲和力的 RNA 适体。筛选出的 RNA 适体特异性地与丹氟沙星结合,但不与四环素结合。RNA 适体的截断至 36 个核苷酸并不影响特异性和亲和力。截断的 RNA 适体特异性地与靶化学物质结合,允许将丹氟沙星与其他氟喹诺酮类药物区分开来。分离出的特异性适体可能成为一种用于快速、经济高效地检测和感测丹氟沙星的潜在试剂,替代包括更昂贵和耗时的高效液相色谱法和液质联用在内的仪器方法。