Department of Chemistry, Capital Normal University, Beijing, 100048, China.
College of Chemical and Environmental Engineering, Shandong University of Science and Technology, Qingdao, 266590, China.
Mikrochim Acta. 2018 Oct 2;185(10):488. doi: 10.1007/s00604-018-3031-z.
An aptamer based assay is described for the colorimetric detection of adenosine. The presence of adenosine triggers the deformation of hairpin DNA oligonucleotide (HP1) containing adenosine aptamer and then hybridizes another unlabeled hairpin DNA oligonucleotide (HP2). This leads to the formation of a double strand with a blunt 3' terminal. After exonuclease III (Exo III)-assisted degradation, the guanine-rich strand (GRS) is released from HP2. Hence, the adenosine-HP1 complex is released to the solution where it can hybridize another HP2 and initiate many cycles of the digestion reaction with the assistance of Exo III. This leads to the generation of a large number of GRS strands after multiple cycles. The GRS stabilize the red AuNPs against aggregation in the presence of potassium ions. If, however, GRS forms a G-quadruplex, it loses its ability to protect gold nanoparticles (AuNPs) from salt-induced AuNP aggregation. Therefore, the color of the solution changes from red to blue which can be visually observed. This colorimetric assay has a 0.13 nM detection limit and a wide linear range that extends from 5 nM to 1 μM. Graphical abstract Schematic presentation of a colorimetric aptamer biosensor for adenosine detection based on DNA cycling amplification and salt-induced aggregation of gold nanoparticles.
一种基于适体的比色法测定腺苷。存在腺苷触发发夹 DNA 寡核苷酸(HP1)中含有腺苷适体的构象变化,然后与另一个未标记的发夹 DNA 寡核苷酸(HP2)杂交。这导致形成具有钝 3'末端的双链体。在核酸外切酶 III(Exo III)辅助降解后,富含鸟嘌呤的链(GRS)从 HP2 中释放出来。因此,腺苷-HP1 复合物被释放到溶液中,在 Exo III 的辅助下可以与另一个 HP2 杂交并引发许多消化反应循环。这导致在多个循环后产生大量的 GRS 链。GRS 在存在钾离子的情况下稳定金纳米粒子(AuNPs)免于聚集。然而,如果 GRS 形成 G-四链体,它就会失去保护金纳米粒子(AuNPs)免受盐诱导的 AuNP 聚集的能力。因此,溶液的颜色从红色变为蓝色,可以通过肉眼观察到。这种比色测定法的检测限为 0.13 nM,线性范围很宽,从 5 nM 延伸到 1 μM。