Department of Pathology, Tianjin Bao Di Hospital, Bao Di Clinical College of Tianjin Medical University, Tianjin, 301800, China.
Key Laboratory of Optic-electric Sensing and Analytical Chemistry for Life Science, MOE, Shandong Key Laboratory of Biochemical Analysis, Key Laboratory of Analytical Chemistry for Life Science in Universities of Shandong, State Key Laboratory Base of Eco-chemical Engineering, Key Laboratory of Rubber-Plastics of Ministry of Education/Shandong Provincial Key Laboratory of Rubber Plastics, College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao, 266042, China.
J Pharm Biomed Anal. 2020 Jul 15;186:113280. doi: 10.1016/j.jpba.2020.113280. Epub 2020 Mar 26.
PtCuCo trimetallic alloys (PtCuCo-TAs) are synthesized by one-step reduction. The chemiluminescence (CL) properties of PtCuCo-TAs are studied systemically. PtCuCo-TAs show good catalyzing for luminol-HO system. A CL platform is developed for the detection of miRNA-21 using PtCuCo-TAs as nanozyme catalyzer. In the CL detection platform, H1 (Hairpin DNA1) is immobilized onto magnetic beads (MBs) firstly. In the presence of miRNA-21, H1 is opened. H2 (Hairpin DNA2) then hybridizes with H1. Meanwhile, a "cleat" in the end of miRNA-21 with a fewer bases complementary is formed to prevent miRNA-21 dissociating from H1. This miRNA-21 hybridizes to another H1. When cpDNA-PtCuCo-TAs which consisted with cDNA (Complementary strand of probe DNA) and pDNA-PtCuCo-TAs (PtCuCo-TAs labeled with probe DNA) are added, the ssDNA region of H1 reacts with the toehold domain of probe DNA and cDNA is released resulting pDNA-PtCuCo-TAs being captured. With this process repeatedly, a lot of pDNA-PtCuCo-TAs are captured onto MBs. After separation and washing, the precipitate and HO are put into the 96-well and luminol solution is injected. The CL signal is produced by PtCuCo-TAs catalyzing luminol-HO system. The amount of miRNA-21 is detected with CL signal. This CL platform performs with limit of detection 0.167 fM and has good selectivity over other RNA.
PtCuCo 三元合金（PtCuCo-TAs）通过一步还原法合成。系统研究了 PtCuCo-TAs 的化学发光（CL）性质。PtCuCo-TAs 对鲁米诺-HO 体系具有良好的催化作用。基于 PtCuCo-TAs 作为纳米酶催化剂，建立了一种用于检测 miRNA-21 的 CL 平台。在 CL 检测平台中，首先将 H1（发夹 DNA1）固定在磁珠（MBs）上。存在 miRNA-21 时，H1 被打开。然后 H2（发夹 DNA2）与 H1 杂交。同时，miRNA-21 形成一个“缺口”，其中碱基互补较少，以防止 miRNA-21 从 H1 上解离。这种 miRNA-21 与另一个 H1 杂交。当加入由 cDNA（探针 DNA 的互补链）和 pDNA-PtCuCo-TAs（标记有探针 DNA 的 PtCuCo-TAs）组成的 cpDNA-PtCuCo-TAs 时，H1 的 ssDNA 区域与探针 DNA 的结合域反应，cDNA 被释放，导致 pDNA-PtCuCo-TAs 被捕获。通过这个过程反复进行，大量的 pDNA-PtCuCo-TAs 被捕获到 MBs 上。分离和洗涤后，将沉淀和 HO 放入 96 孔板中，并注入鲁米诺溶液。CL 信号由 PtCuCo-TAs 催化鲁米诺-HO 体系产生。通过 CL 信号检测 miRNA-21 的含量。该 CL 平台的检测限为 0.167 fM，对其他 RNA 具有良好的选择性。
