Food and Environment Research Agency, Sand Hutton, York YO41 1LZ North Yorkshire, United Kingdom.
Anal Chem. 2010 Apr 1;82(7):2652-60. doi: 10.1021/ac902226v.
A robust screening assay employing solid phase extraction (SPE) followed by a novel aptamer-based procedure is presented for the rapid detection and semiquantitation of the triphenylmethane dye, Malachite Green (MG) and its primary metabolite Leucomalachite Green (LMG) in fish tissue. To the authors' knowledge, this is the first reported use of an RNA aptamer for the development of a diagnostic assay for the detection of chemical residues in food. The aptamer based screening assay is found to be highly specific for MG; but has negligible affinity for the LMG metabolite. However, because the LMG metabolite is lipophilic and known to be highly persistent in tissues, an oxidation step has been incorporated within the sample cleanup procedure to ensure that all LMG residues are converted to MG prior to measurement. This article provides evidence that an oligonucleotide aptamer can be used as an alternative recognition element to conventional antibodies with application to the detection of residues in food. Furthermore, this finding has the future potential to reduce the number of animals currently being used in the production of antibodies for immunodiagnostic kits.
一种基于固相萃取（SPE）和新型适体的可靠筛选方法，用于快速检测和半定量鱼肉组织中的三苯甲烷染料孔雀石绿（MG）及其主要代谢产物隐色孔雀石绿（LMG）。据作者所知，这是首次报道使用 RNA 适体开发用于检测食品中化学残留物的诊断检测方法。基于适体的筛选检测方法对 MG 具有高度特异性；但对 LMG 代谢物的亲和力可以忽略不计。然而，由于 LMG 代谢物具有亲脂性，并且已知在组织中高度持久，因此在样品净化过程中加入了氧化步骤，以确保在测量之前将所有 LMG 残留转化为 MG。本文提供的证据表明，寡核苷酸适体可以用作替代传统抗体的识别元件，应用于食品中残留物的检测。此外，这一发现具有减少目前用于免疫诊断试剂盒生产的动物数量的未来潜力。
