Selection, identification and application of a DNA aptamer against Staphylococcus aureus enterotoxin A
Selection, identification and application of a DNA aptamer against Staphylococcus aureus enterotoxin A
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DOI：
10.1039/c3ay41576g
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发表时间：
2014-01-01
影响因子：
3.1
通讯作者：
Wang, Zhouping
中科院分区：
文献类型：
--
作者：
Huang, Yukun;Chen, Xiujuan;Wang, Zhouping
近年来，由金黄色葡萄球菌肠毒素(SES)污染的食物引起的葡萄球菌食物中毒(SFP)已成为世界范围内最常见的食源性疾病之一。因此，快速、灵敏和可靠的检测方法对于常规的SE观察至关重要。然而，目前的检测方法主要是抗体依赖的，而且检测过程中新鲜抗体的制备和不稳定限制了方法的发展。在本研究中，通过基于磁分离技术的十二轮筛选过程，在体外获得了与金黄色葡萄球菌肠毒素A(SEA)高亲和力和选择性的适配子，其解离常数(K-d)值低至48.57+/-6.52 nm。优化的适体A15成功地用于食品样品中SEA的荧光生物检测，检出限为8.7×10~(-3)µg·mL~(-1)。基于这项研究，所选择的适配子有望成为新的分子识别元件，可用于海洋探测的创新生物传感器。
In recent years, staphylococcal food poisoning (SFP) caused by the ingestion of food contaminated with Staphylococcus aureus enterotoxins (SEs) has been one of the most common foodborne illnesses worldwide. As a result, rapid, sensitive and reliable detection methods are crucial for routine observations of SEs. However, current detection methods are primarily antibody-dependent, and the development of methods is limited by the preparation and instability of fresh antibodies during testing. In this study, aptamers that bind to Staphylococcus aureus enterotoxin A (SEA) with high affinity and selectivity were generated in vitro by a twelve-round selection process based on magnetic separation technology with a dissociation constant (K-d) value as low as 48.57 +/- 6.52 nM. The optimal aptamer A15 was successfully used in the fluorescent bioassay to detect SEA in the food sample with a detection limit of 8.7 x 10(-3) mu g mL(-1). Based on this study, the selected aptamers can be expected to be new molecular recognition elements that can be used in innovative biosensors for SEA detections.