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.

禁用的抗病毒兽药金刚烷胺（AMD）仍在滥用。这项研究开发了一种新型的酶联免疫吸附测定法，用于AMD的比色检测，涉及DNA杂交反应和非交联金纳米颗粒（AuNPs）聚集。因此，引物1–AuNPs–抗AMD单克隆抗体（mAb）可以被ELISA孔中的AMD人工抗原捕获。与引物1互补配对的引物2最终被加入到ELISA孔中。杂交反应后，将上清液中的游离引物2与AuNPs和NaCl混合，并诱导AuNPs快速变色。样品中缺乏AMD导致捕获了大量的Primer 1–AuNPs–mAb复合物，并在上清液中捕获了有限的游离Primer 2。添加NaCl后，AuNPs的颜色在有限的Primer 2情况下变为蓝色。