Department of Chemistry, Institute of Interdisciplinary Convergence Research, Research Institute of Halal Industrialization Technology, Chung-Ang University, 84 Heukseok-ro, Donjak-gu, Seoul 06974, Republic of Korea.
Department of Pharmaceutical Engineering, Daegu Haany University, Gyeongsan 38610, Republic of Korea.
Biosens Bioelectron. 2018 Dec 15;121:111-117. doi: 10.1016/j.bios.2018.08.068. Epub 2018 Aug 28.
Herein, a straightforward and highly specific dot-blot immunoassay was successfully developed for the detection of Mycobacterium tuberculosis antigen (10 kDa culture filtrate protein, CFP-10) via the formation of copper nanoshell on the gold nanoparticles (AuNPs) surface. The principle of dot-blot immunoassay was based on the reduction of Cu ion on the GBP-CFP10G2-AuNPs conjugates, which has gold binding and antigen binding affinities, simultaneously, favouring to appear red dot that can be observed with naked-eye. The dot intensity is proportional to the concentration of tuberculosis antigen CFP-10, which offers a detection limit of 7.6 pg/mL. The analytical performance of GBP-CFP10G2-AuNPs-copper nanoshell dot-blot was superior than that of conventional silver nanoshell. This method was successfully applied to identify the CFP-10 antigen in the clinical urine sample with high sensitivity, specificity, and minimized sample preparation steps. This method exhibits great application potential in the field of nanomedical science for highly reliable point-of-care detection of CFP-10 antigen in real samples to early diagnosis of tuberculosis.
本文通过在金纳米粒子（AuNPs）表面形成铜纳米壳，成功开发了一种用于检测结核分枝杆菌抗原（10kDa 培养滤液蛋白，CFP-10）的简单且高度特异的斑点印迹免疫分析方法。斑点印迹免疫分析的原理基于在 GBP-CFP10G2-AuNPs 缀合物上 Cu 离子的还原，该缀合物具有金结合和抗原结合亲和力，同时有利于出现可以用肉眼观察到的红点。斑点强度与结核抗原 CFP-10 的浓度成正比，检测限为 7.6pg/mL。GBP-CFP10G2-AuNPs-铜纳米壳斑点印迹的分析性能优于传统的银纳米壳。该方法成功应用于临床尿液样本中 CFP-10 抗原的高灵敏度、特异性和最小化样本制备步骤的鉴定。该方法在纳米医学领域具有很大的应用潜力，可用于在实际样本中进行高度可靠的 CFP-10 抗原的即时检测，以实现结核病的早期诊断。
