Faculty of Material Science and Chemical Engineering, Ningbo University, Ningbo 315211, China; Key Laboratory of Animal Protein Food Processing Technology of Zhejiang Province, Ningbo University, Ningbo 315211, China.
Faculty of Material Science and Chemical Engineering, Ningbo University, Ningbo 315211, China.
Biosens Bioelectron. 2019 Apr 1;130:139-146. doi: 10.1016/j.bios.2019.01.044. Epub 2019 Jan 29.
Microchip electrophoresis (MCE) was a good available method for high-throughput and rapid detecting chemical pollutants in food samples. However, many of the reported MCE assays involve complex design of microchip, laborious operation and poor universality which limited its promotion in multiple antibiotics' detection. Herein, a multiplexed aptasensor was developed based on a universal double-T type microchip to one-step and simultaneously detect several antibiotics within 3 min using chloramphenicol (CAP) and kanamycin (Kana) as representatives. Besides, a novel stir-bar assisted DNA multi-arm junctions recycling (MAJR) strategy was designed for transducing and amplifying the signal. The brief detection mechanism was as following: the added CAP and Kana can specifically react with their aptamer probes on the stir-bar and produce different single-stranded DNA primer, respectively. Afterwards, the primers can trigger MAJR to form a lot of three- and four-arm DNA junctions corresponding to different targets. The DNA multi-arm junctions can be easily separated and detected by MCE for quantification. Moreover, the stir-bar can facilitate phase separation and obviously eliminate matrix interference in food. The assay was successfully applied in milk and fish samples, showing excellent selectivity and sensitivity with a detection limits of 0.52 pg mL CAP and 0.41 pg mL Kana (S/N = 3). Thus, the assay holds a great potential application for screening of antibiotics in food.
微芯片电泳 (MCE) 是一种高通量、快速检测食品样品中化学污染物的有效方法。然而，许多报道的 MCE 分析方法涉及微芯片的复杂设计、繁琐的操作和较差的通用性，这限制了其在多种抗生素检测中的推广。本文基于通用的双 T 型微芯片，开发了一种多重适体传感器，可在 3 分钟内一步同时检测几种抗生素，以氯霉素 (CAP) 和卡那霉素 (Kana) 为代表。此外，设计了一种新颖的搅拌棒辅助 DNA 多臂 junctions recycling (MAJR) 策略来转换和放大信号。简要的检测机制如下：添加的 CAP 和 Kana 可以与搅拌棒上的适体探针特异性反应，分别产生不同的单链 DNA 引物。然后，这些引物可以触发 MAJR 形成与不同靶标相对应的大量三臂和四臂 DNA 连接物。DNA 多臂连接物可以通过 MCE 轻松分离和检测，用于定量。此外，搅拌棒可以促进相分离，并明显消除食品中的基质干扰。该方法成功应用于牛奶和鱼肉样品，表现出优异的选择性和灵敏度，对 CAP 的检测限为 0.52 pg mL，对 Kana 的检测限为 0.41 pg mL (S/N=3)。因此，该方法在食品中抗生素的筛选方面具有很大的应用潜力。
