A competitive fluorescent aptasensor for okadaic acid detection assisted by rolling circle amplification
A competitive fluorescent aptasensor for okadaic acid detection assisted by rolling circle amplification
复制标题
滚环扩增辅助检测冈田酸的竞争性荧光适体传感器
DOI：
10.1007/s00604-017-2293-1
复制
发表时间：
2017-05
影响因子：
5.7
通讯作者：
Wang Zhouping
中科院分区：
文献类型：
--
作者：
Gu Huajie;Hao Liling;Duan Nuo;Wu Shijia;Xia Yu;Ma Xiaoyuan;Wang Zhouping
冈田酸（Okadaic acid，OA）是一种低分子量的海洋贝类毒素，可引起贝类中毒。由于其频繁发生，OA已成为严重威胁人类健康和水产品加工业的一个重要因素。作者描述了一种基于滚环扩增（RCA）的OA竞争性荧光团连接适体测定法。其由以下步骤组成：（a）通过将OA适体固定在微孔板的表面上来修饰微孔板的威尔斯孔;（B）使适体与适体-互补序列-引物复合物杂交;（c）进行RCA反应;（d）加入FAM标记的信号探针。OA与检测探针竞争固定的适体。竞争反应发生后，除去含有释放的检测探针的上清液，然后用酶标仪读数。与信号与OA浓度负相关的竞争性测定不同，该方法在荧光强度和OA浓度之间具有正相关性。优化后的方法具有较低的检测限（1 pg·mL−1）和较宽的线性范围（1 pg·mL− 1 ~ 100 ng·mL−1），这是由于RCA信号放大的缘故。该方法特异性强，重复性好，回收率高，可用于海产品中OA的检测。图解摘要：采用荧光团连接的适体辅助滚环扩增（RCA）的冈田酸微孔板检测方法的原理图。信号强度与OA的浓度成正比。还进行了无RCA的试验，并与进行了比较。
Okadaic acid (OA) is a low molecular weight marine toxin from shellfish which causes diarrheic shellfish poisoning (DSP). Due to its frequent occurrence, OA has become a serious threat to human health and seafood industry. The authors describe a competitive fluorophore-linked aptamer assay for OA that is based on rolling circle amplification (RCA). It consists of the following steps: (a) The wells of a microplate are modified by fixing the OA aptamer on their surface; (b) The aptamer is hybridized with an aptamer-complementary sequence-primer complex; (c) the RCA reaction is performed; (d) the FAM labelled signal probe is added. OA competes with the detection probe for the immobilized aptamer. After the competitive reaction has occurred, the supernatants containing released detection probes are removed and then read with a microplate reader. This method, unlike in competitive assays where the signals negatively correlate with OA concentrations, has a positive correlation between fluorescence intensity and OA concentration. The optimized assay has a lower detection limit (1 pg·mL−1) and a wider linear range (from 1 pg·mL−1to 100 ng·mL−1) owning to signal amplification via RCA. It also is highly specific, repeatable, has good recovery and can be used to detect OA in seafood.Graphical abstractSchematic of a microplate assay for okadaic acid assisted by rolling circle amplification (RCA) and using a fluorophore-linked aptamer. The signal intensities are directly proportional to the concentrations of OA. The assay without RCA was also performed and compared to.
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影响因子：
5.4
作者：
Valdiglesias V;Prego-Faraldo MV;Pásaro E;Méndez J;Laffon B
通讯作者：
Laffon B
DOI：
--
发表时间：
2015
期刊：
Journal of Food Safety and Quality
影响因子：
--
作者：
Wu Zhiqiang;W. Bencheng;Su YanBo;Liu Yi
通讯作者：
Wu Zhiqiang;W. Bencheng;Su YanBo;Liu Yi
影响因子：
4.1
作者：
D. Garibo;E. Devic;J. Marty;J. Diogène;I. Unzueta;M. Blázquez;M. Campàs
通讯作者：
D. Garibo;E. Devic;J. Marty;J. Diogène;I. Unzueta;M. Blázquez;M. Campàs
影响因子：
8.4
作者：
Zou, Ling;Wang, Qin;Wang, Ping
通讯作者：
Wang, Ping
影响因子：
6.1
作者：
Liu, Biing-Hui;Hung, Chun-Tse;Yu, Feng-Yih
通讯作者：
Yu, Feng-Yih