Fluorometric determination of HIV DNA using molybdenum disulfide nanosheets and exonuclease III-assisted amplification
Fluorometric determination of HIV DNA using molybdenum disulfide nanosheets and exonuclease III-assisted amplification
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使用二硫化钼纳米片和核酸外切酶 III 辅助扩增荧光测定 HIV DNA
DOI：
10.1007/s00604-019-3368-y
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发表时间：
2019
影响因子：
5.7
通讯作者：
Wen Yanli
中科院分区：
文献类型：
--
作者：
Wang Lele;Dong Lianhua;Liu Gang;Shen Xizhong;Wang Jing;Zhu Changfeng;Ding Min;Wen Yanli
本文介绍了一种简便、灵敏的荧光法测定HIV DNA。它利用了MoS 2纳米片对长ssDNA与短寡核苷酸片段的亲和力的强烈差异。此外，有效的信号放大是通过核酸外切酶III辅助的目标再循环来实现的。当吸附在MoS 2纳米片上时，FAM标记的ssDNA探针（FP）的荧光被淬灭。然而，在HIV DNA存在的情况下，FP与靶标杂交形成双链体。结果，双链体中的FP将被Exo III逐步水解成短片段，并且由于短片段对MoS 2纳米片具有低亲和力，因此保留了荧光信号。通过使用Exo III辅助的靶循环扩增，检测灵敏度大大提高。该传感器可以检测浓度低至5.3 pM的DNA（S/N比为3），分析范围从0.01 nM扩展到10 nM。该检测方法简单，灵敏，特异性强，可以想象，代表了一个有价值的工具，在临床研究相关的HIV。图形abstraction的荧光测定的基础上二硫化钼纳米片和Exo III的HIV DNA的示意图。当荧光标记的ssDNA探针与靶标杂交形成双链体时，产生Exo III辅助的靶标再循环扩增。该方法可以检测低至5.3 pM的HIV DNA。
A convenient and ultrasensitive fluorometric method is described for the determination of HIV DNA. It exploits the strong difference in the affinities of MoS2nanosheets for long ssDNA versus short oligonucleotide fragments. In addition, efficient signal amplification is accomplished by exonuclease III-assisted target recycling. When absorbed on the MoS2nanosheets, the fluorescence of the FAM-labeled ssDNA probe (FP) is quenched. However, in the presence of HIV DNA, the FP hybridizes with target to form a duplex. As a result, the FP in the duplex will be stepwise hydrolyzed into short fragments by Exo III, and the fluorescence signal thus is retained because short fragments have low affinity for the MoS2nanosheets. By using the Exo III-assisted target recycling amplification, the detection sensitivity is strongly improved. The sensor can detect DNA in a concentration as low as 5.3 pM (at an S/N ratio of 3), and the analytical range extends from 0.01 nM to 10 nM. The assay is simple, sensitive and specific, and conceivably represents a valuable tool in clinical studies related to the HIV.Graphical abstractSchematic presentation of fluorometric determination of HIV DNA based on molybdenum disulfide nanosheets and Exo III. When the fluorescence-tagged ssDNA probe hybridized with target to form a duplex, the Exo III-assisted target recycling amplification is generated. The method can detect as low as 5.3 pM HIV DNA.