Zhang Jingjing, Han Jialun, Feng Shehong, Niu Chaoqun, Liu Chen, Du Jie, Chen Yong
State Key Laboratory of Marine Resource Utilization in South China Sea, College of Information Science & Technology, Hainan University, Haikou 570228, China.
College of Materials & Chemistry Engineering, Hainan University, Haikou 570228, China.
Materials (Basel). 2018 Nov 29;11(12):2408. doi: 10.3390/ma11122408.
In this study, a target recycled amplification, background signal suppression, label-free fluorescent, enzyme-free deoxyribonucleic acid (DNA) machine was developed for the detection of adenosine triphosphate (ATP) in human urine. ATP and DNA fuel strands (FS) were found to trigger the operation of the DNA machine and lead to the cyclic multiplexing of ATP and the release of single stranded (SS) DNA. Double-stranded DNA (dsDNA) was formed on graphene oxide (GO) from the combination of SS DNA and complementary strands (CS'). These double strands then detached from the surface of the GO and in the process interacted with PicoGreen dye resulting in amplifying fluorescence intensity. The results revealed that the detection range of the DNA machine is from 100 to 600 nM (R² = 0.99108) with a limit of detection (LOD) of 127.9 pM. A DNA machine circuit and AND-NOT-AND-OR logic gates were successfully constructed, and the strategy was used to detect ATP in human urine. With the advantage of target recycling amplification and GO suppressing background signal without fluorescent label and enzyme, this developed strategy has great potential for sensitive detection of different proteins and small molecules.
在本研究中，开发了一种用于检测人尿中三磷酸腺苷（ATP）的目标循环扩增、背景信号抑制、无标记荧光、无酶脱氧核糖核酸（DNA）机器。发现ATP和DNA燃料链（FS）触发DNA机器的运行，并导致ATP的循环复用和单链（SS）DNA的释放。SS DNA与互补链（CS'）结合在氧化石墨烯（GO）上形成双链DNA（dsDNA）。这些双链随后从GO表面脱离，在此过程中与PicoGreen染料相互作用，导致荧光强度放大。结果表明，DNA机器的检测范围为100至600 nM（R² = 0.99108），检测限（LOD）为127.9 pM。成功构建了DNA机器电路和与非与或逻辑门，并将该策略用于检测人尿中的ATP。凭借目标循环扩增以及GO在无荧光标记和酶的情况下抑制背景信号的优势，这种开发的策略在灵敏检测不同蛋白质和小分子方面具有巨大潜力。
