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Selection, identification and application of a DNA aptamer against Staphylococcus aureus enterotoxin A

机译:抗金黄色葡萄球菌肠毒素A的DNA适体的选择,鉴定和应用

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摘要

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 foodbome 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 × 10~(-3) μg 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.
机译:近年来,由于摄入被金黄色葡萄球菌肠毒素(SEs)污染的食物而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物病。因此,快速,灵敏和可靠的检测方法对于SE的常规观察至关重要。然而,当前的检测方法主要是抗体依赖性的,并且方法的发展受到新鲜抗体的制备和测试期间的不稳定性的限制。在这项研究中,通过基于磁分离技术的十二轮选择过程,以离解常数(K_d)值低至48.57±6.52的体外十二轮选择过程,以高亲和力和选择性结合了与金黄色葡萄球菌肠毒素A(SEA)结合的适体。 nM,最佳适体A15成功用于荧光生物测定中,以8.7×10〜(-3)μgmL〜(-1)的检测限检测食品样品中的SEA。根据这项研究,可以预期所选的适体是新的分子识别元件,可用于SEA检测的创新生物传感器。

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