State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, China.
Jiangxi Province Center for Disease Control and Prevention, Nanchang 330047, China.
Food Chem. 2018 Aug 15;257:382-387. doi: 10.1016/j.foodchem.2018.03.033. Epub 2018 Mar 9.
Amantadine (AMD), a banned antiviral veterinary drug, is still being abused. This study developed a novel enzyme linked immunosorbent assay for the colorimetric detection of AMD involving DNA hybridization reaction and non-crosslinking gold nanoparticles (AuNPs) aggregation. Accordingly, the Primer 1-AuNPs-anti-AMD monoclonal antibody (mAb) could be captured by AMD artificial antigen on ELISA wells. Primer 2, which was complementary paired to Primer 1, was eventually added into the ELISA wells. After the hybridization reaction, the free Primer 2 in the supernatant was mixed with AuNPs and NaCl and induced a rapid color change of AuNPs. The lack of AMD in the sample resulted in capturing a substantial Primer 1-AuNPs-mAb complex and limited free Primer 2 in the supernatant. After adding NaCl, the color of AuNPs turned blue with limited Primer 2. This simple and visualized novel method had good sensitivity (0.033 μM) and exhibited a potential application for AMD screening on site.
金刚烷胺（AMD）是一种已被禁用的抗病毒兽用药物，但仍被滥用。本研究开发了一种新型酶联免疫吸附测定法（ELISA），用于比色检测 AMD，涉及 DNA 杂交反应和非交联金纳米粒子（AuNPs）聚集。因此，Primer 1-AuNPs-抗 AMD 单克隆抗体（mAb）可被 ELISA 孔中的 AMD 人工抗原捕获。随后，互补配对的 Primer 2 被加入到 ELISA 孔中。杂交反应后，上清液中的游离 Primer 2 与 AuNPs 和 NaCl 混合，诱导 AuNPs 快速变色。样品中缺乏 AMD 会导致大量的 Primer 1-AuNPs-mAb 复合物被捕获，上清液中游离的 Primer 2 受到限制。加入 NaCl 后，由于 Primer 2 有限，AuNPs 的颜色变为蓝色。这种简单可视化的新型方法具有良好的灵敏度（0.033 μM），并有望在现场用于 AMD 的筛选。
