Smiley Sally, Derosa Maria, Blais Burton
Ottawa Laboratory (Carling), Canadian Food Inspection Agency, Ottawa, ON, Canada.
J Nucleic Acids. 2013;2013:936542. doi: 10.1155/2013/936542. Epub 2013 Oct 30.
A simple dot blot immunoenzymatic assay system was developed using polyester cloth coated with an oligo-DNA aptamer to provide a high-affinity macroporous surface for the efficient capture of a model protein analyte (thrombin) in complex sample matrices such as foods. Bound thrombin was detected immunoenzymatically using a peroxidase-linked antithrombin antibody and a chromogenic substrate. A unique feature of this approach, which we have termed "aptablot," is the facile immobilization of DNA aptamers on the polyester surface by cross-linking with a brief exposure to ultraviolet light, and the simple assay format obviating the need for specialized instruments. The assay principle described herein should be broadly applicable to many situations where analytes must be detected in complex samples, with the main limiting factor being the availability of suitable DNA aptamers.
开发了一种简单的斑点印迹免疫酶分析系统，该系统使用涂有寡聚DNA适配体的聚酯布，以提供一个高亲和力的大孔表面，用于在诸如食品等复杂样品基质中有效捕获模型蛋白质分析物（凝血酶）。使用过氧化物酶标记的抗凝血酶抗体和显色底物通过免疫酶法检测结合的凝血酶。我们将这种方法称为“适配体印迹法”，其独特之处在于通过短暂暴露于紫外线下进行交联，可轻松将DNA适配体固定在聚酯表面，并且简单的分析形式无需专门的仪器。本文所述的分析原理应广泛适用于许多需要在复杂样品中检测分析物的情况，主要限制因素是合适的DNA适配体的可用性。
