Aptamer based ultrasensitive determination of the beta-adrenergic agonist ractopamine using PicoGreen as a fluorescent DNA probe

文献类型： 外文期刊

第一作者： Zhu, Chao

作者： Zhu, Chao;Zhang, Guilan;Huang, Yafei;Yan, Jiao;Chen, Ailiang;Zhu, Chao;Zhang, Guilan;Huang, Yafei;Yan, Jiao;Chen, Ailiang;Huang, Yafei;Yan, Jiao

作者机构：

关键词： Aptamer;Ractopamine;PicoGreen;Swine urine;dsDNA;DNA probe;Fluorescence assay

期刊名称：MICROCHIMICA ACTA （ 影响因子：5.833； 五年影响因子：5.357 ）

ISSN：

年卷期：

页码：

收录情况： SCI

摘要： The authors describe an aptamer based assay for determination of ractopamine (RAC) by using PicoGreen (PG) as a fluorescent probe specific for dsDNA. In the absence of RAC, the aptamer forms a duplex structure with a complementary sequence that results in enhanced PG fluorescence. Upon binding to RAC, the aptamer undergoes a structural switch. This reduces the number of DNA duplexes formed and causes a reduction of fluorescence intensity of PG as measured at excitation/emission wavelengths of 480/520 nm. Under optimized conditions, the dynamic calibration plot covers the 50 pM to 50 mu M concentration range, with a 50 pM detection limit. This meets the safety supervision regulations of the European Commission in terms of residue limits of RAC in food. The method displays high selectivity over other beta-adrenergic agonists including clenbuterol, dopamine and salbutamol. The assay was successfully applied to samples of swine urine at spiking levels of 7.4 nM, 22.2 nM and 37 nM. Average recoveries ranged from 95 to 110%, with an RSD of < 1.5%. The method is expected to represent a promising tool for simple, rapid and sensitive on-site detection of RAC in animal products.

分类号： O65`O655

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