Aptameric biosensor for the sensitive detection of major shrimp allergen, tropomyosin
Aptameric biosensor for the sensitive detection of major shrimp allergen, tropomyosin
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DOI：
10.1016/j.foodchem.2019.126133
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发表时间：
2020-06-01
期刊：
影响因子：
8.8
通讯作者：
Zourob, Mohammed
中科院分区：
文献类型：
--
作者：
Chinnappan, Raja;Rahamn, Anas Abdel;Zourob, Mohammed
开发一种灵敏、快速的方法来检测各种食品基质中的过敏原对于帮助患者控制过敏至关重要。最常用的过敏原检测方法是基于免疫测定、PCR 和质谱法。然而，所有这些都非常复杂且耗时。在此，开发了一种用于检测主要虾过敏原原肌球蛋白（TM）的适体生物传感器。氧化石墨烯（GO）被用作筛选高亲和力靶标结合所需的最小长度适体序列的平台。标记 GO 的荧光素染料通过 p 堆积相互作用猝灭截短的适体。添加TM后，由于适体与GO的竞争性结合，荧光恢复。发现其中一种截短的适体与 TM 的结合亲和力 (30 nM) 是全长适体 (124 nM) 的四倍，检测限 (LOD) 为 2 nM。基于适配体的传感器展示了 30 分钟内对 TM 的灵敏、选择性和特异性检测。使用 TM 加标鸡汤确认了传感器的性能，从而实现了高回收率（类似于 97 +/- 10%）。 GO与标记适体传感器平台的结合显示出食品中TM的快速检测，与其他方法相比非常灵敏、特异并且在高通量应用中表现良好。
The development of a sensitive and rapid detection approach for allergens in various food matrices is essential to assist patients in managing their allergies. The most common methods used for allergen detection are based on immunoassays, PCR and mass spectrometry. However, all of them are very complex and time-consuming. Herein, an aptamer biosensor for the detection of the major shrimp allergen tropomyosin (TM) was developed. Graphene oxide (GO) was used as a platform for screening of the minimal-length aptamer sequence required for high-affinity target binding. A fluorescein dye labeled GO quenches the truncated aptamer by p-stacking interactions. After the addition of TM, the fluorescence was restored due to the competitive binding of the aptamer to GO. One of the truncated aptamers was found to bind to TM with four-fold higher affinity (30 nM) compared to the full-length aptamer (124 nM), with a limit of detection (LOD) of 2 nM. The aptamer-based sensor demonstrates the sensitive, selective, and specific detection of TM in 30 min. The performance of the sensor was confirmed using TM spiked chicken soup, resulting in a high percentage recovery (similar to 97 +/- 10%). The association of GO and labelled aptamer sensor platform has shown the rapid detection of TM in food, which is compared to other methods very sensitive, specific and performs in high throughput application.