Zhao Yuanfang, Zhao Jingru, Jin Tian, Sun Shuqing, Liu Wenlan, Tan Ying
Institute of Optical Imaging and Sensing, Shenzhen Key Laboratory for Minimal Invasive Medical Technologies, Graduate School at Shenzhen, Tsinghua University Shenzhen 518055 P. R. China
Open FIESTA, Graduate School at Shenzhen, Tsinghua University Shenzhen 518055 P. R. China.
RSC Adv. 2019 Oct 24;9(59):34293-34298. doi: 10.1039/c9ra06203c. eCollection 2019 Oct 23.
C-reactive protein (CRP) is a crucial clinical biomarker for inflammatory and cardiovascular diseases. Therefore, the sensitive, selective and convenient detection of CRP is of great significance. Using gold nanoparticles (AuNPs) and combining the specific interaction between an aptamer and CRP, we developed a simple and convenient assay for CRP detection. The aptamer-based probe was fabricated through the hybridization of CRP-aptamer immobilized on magnetic beads (MBs) to a short complementary DNA (cDNA) chain attached to AuNPs to form a MB-Aptamer-AuNP sandwich structure. Upon the addition of CRP, aptamer-cDNA dehybridization occurred due to the strong interaction between CRP and the aptamer, resulting in the release of AuNPs, which were subjected to DFM imaging and subsequently counted using the MATLAB program. The number of AuNPs was therefore positively correlated to the concentration of CRP and a detection limit as low as 2.71 nM was achieved. The current approach could also exclude the disturbance of other proteins, including thrombin, IgG, Lys and BSA. In addition, the concentration of CRP detected was in good agreement with the amount cast in bovine and mouse serum, indicating that the proposed probe is robust and accurate, and it is very promising for practical applications where CRP detection is necessary. The current strategy is also promising for the detection of other proteins where a suitable aptamer is selected.
C反应蛋白(CRP)是炎症和心血管疾病的关键临床生物标志物。因此,灵敏、选择性好且便捷地检测CRP具有重要意义。利用金纳米颗粒(AuNPs)并结合适体与CRP之间的特异性相互作用,我们开发了一种简单便捷的CRP检测方法。基于适体的探针通过将固定在磁珠(MBs)上的CRP适体与连接到AuNPs的短互补DNA(cDNA)链杂交来构建,形成MB-适体-AuNP夹心结构。加入CRP后,由于CRP与适体之间的强相互作用,适体-cDNA发生解杂交,导致AuNPs释放,对其进行暗场显微镜(DFM)成像,随后使用MATLAB程序进行计数。因此,AuNPs的数量与CRP浓度呈正相关,检测限低至2.71 nM。当前方法还可以排除其他蛋白质(包括凝血酶、IgG、赖氨酸和牛血清白蛋白)的干扰。此外,检测到的CRP浓度与牛血清和小鼠血清中添加的量高度一致,表明所提出的探针可靠且准确,在需要检测CRP的实际应用中非常有前景。当前策略对于选择合适适体检测其他蛋白质也很有前景。