Chengdu Institute of Biology, Chinese Academy of Sciences, No.9, Section 4, South Renmin Road, Chengdu, Sichuan, China; University of Chinese Academy of Sciences, Beijing, China.
Chengdu Institute of Biology, Chinese Academy of Sciences, No.9, Section 4, South Renmin Road, Chengdu, Sichuan, China.
J Chromatogr A. 2018 Sep 28;1569:222-228. doi: 10.1016/j.chroma.2018.07.051. Epub 2018 Jul 17.
An aptamer-based microchip capillary electrophoresis coupled with laser induced fluorescence (MCE-LIF) detection method for fast determination of Aflatoxin B (AFB) and Ochratoxin A (OTA) was developed. Aptamers that are specific to these two mycotoxins were first hybridized with their aptamer complementary oligonucleotides. The double strand DNA that comes in contact with mycotoxin-containing environment would be unwound into separate aptamer-mycotoxin complex and aptamer complementary single strand. Different types of oligonucleotides can be separated in MCE and detected under the aid of fluorescent dye SYBR gold in LIF detection unit. Under the optimal conditions, on-chip aptamer-mycotoxin conjugates analysis was achieved within 3 min with extremely low LODs (0.026 ng/mL for AFB and 0.021 ng/mL for OTA). Specificity study indicated that other major mycotoxins would not cross-react with these two aptamers, demonstrating the good selectivity of the proposed method. Quantification of trace AFB and OTA in real food samples was carried out and satisfactory recoveries were obtained. It is demonstrated that this method is fast, facile and specific for Simultaneous determination of trace AFB and OTA from foodstuffs.
建立了基于适体的微芯片毛细管电泳结合激光诱导荧光（MCE-LIF）检测方法，用于快速测定黄曲霉毒素 B（AFB）和赭曲霉毒素 A（OTA）。首先将与这两种真菌毒素特异性结合的适体与适体互补寡核苷酸杂交。与含真菌毒素环境接触的双链 DNA 会被解开成单独的适体-真菌毒素复合物和适体互补单链。在 MCE 中可以分离不同类型的寡核苷酸，并在 LIF 检测单元中荧光染料 SYBR 金的辅助下进行检测。在最佳条件下，芯片上的适体-真菌毒素结合物分析在 3 分钟内完成，LOD 非常低（AFB 为 0.026ng/mL，OTA 为 0.021ng/mL）。特异性研究表明，其他主要真菌毒素不会与这两种适体发生交叉反应，表明该方法具有良好的选择性。对实际食品样品中的痕量 AFB 和 OTA 进行了定量检测，获得了令人满意的回收率。结果表明，该方法快速、简便、特异性强，可用于同时测定食品中的痕量 AFB 和 OTA。
