Gonyautoxin 1/4 aptamers with high-affinity and high-specificity: From efficient selection to aptasensor application
Gonyautoxin 1/4 aptamers with high-affinity and high-specificity: From efficient selection to aptasensor application
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DOI：
10.1016/j.bios.2016.01.032
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发表时间：
2016-05-15
影响因子：
12.6
通讯作者：
Wang, Lianghua
中科院分区：
文献类型：
--
作者：
Gao, Shunxiang;Hu, Bo;Wang, Lianghua
鬼龙鱼毒素1/4(GTX1/4)是一种对公众健康有重大影响的强效海洋神经毒素。然而，与目前应用的麻风性贝毒素GTX1/4检测方法有关的伦理问题和技术缺陷正在推动进一步的研究，以开发监管监测系统中的适当替代品。这项工作首次成功地筛选、优化和鉴定了一个与GTX1/4高亲和力和特异性的适体。与典型的MB-SELEX相比，GO-SELEX作为一种先进的筛选技术，在开发小分子适配子方面具有显著的优势。此外，我们还截短了GTX1/4适配子，得到了适体核心序列，其K-d值较高，为17.7 nM。利用适体GO18-T-d构建了检测GTX1/4的无标记实时光学BLI适体传感器。该适体传感器的检测范围为0.2~200 ng/mLGTX1/4(线性范围为0.2~90 ng/mLGTX1/4)，检测下限为50pg/mL。此外，该适配子传感器对GTX1/4具有高度的特异性，与其他海洋毒素无交叉反应。将该适配子传感器用于贝类样品中GTX1/4的检测，具有良好的重现性和稳定性。我们相信，这种新型的适配传感器为海洋生物毒素GTX1/4的快速检测提供了一种有前途的传统分析方法的替代方法。(C)2016爱思唯尔公司版权所有。
Gonyautoxin 1/4 (GTX1/4) are potent marine neurotoxins with significant public health impact. However, the ethical issues and technical defects associated with the currently applied detection methods for paralytic shellfish toxin GTX1/4 are pressing further studies to develop suitable alternatives in a regulatory monitoring system. This work describes the first successful selection, optimization, and characterization of an aptamer that bind with high affinity and specificity to GTX1/4. Compared to the typical MB-SELEX, GO-SELEX, an advanced screening technology, has significant advantages for small molecular aptamer development. Furthermore, we truncated GTX1/4 aptamer and obtained the aptamer core sequence with a higher K-d of 17.7 nM. The aptamer GO18-T-d was then used to construct a label-free and real-time optical BLI aptasensor for the detection of GTX1/4. The aptasensor showed a broad detection range from 0.2 to 200 ng/mL GTX1/4 (linear range from 0.2 to 90 ng/mL), with a low detection limit of 50 pg/mL. Moreover, the aptasensor exhibited a high degree of specificity for GTX1/4 and no cross reactivity to other marine toxins. The aptasensor was then applied to the detection of GTX1/4 in spiked shellfish samples and showed a good reproducibility and stability. We believe that this novel aptasensor offers a promising alternative to traditional analytical methods for the rapid detection of the marine biotoxin GTX1/4. (C) 2016 Elsevier B.V. All rights reserved.