Detection of HIV/HCV virus DNA with homogeneous DNA machine-triggered in situ formation of silver nanoclusters
Detection of HIV/HCV virus DNA with homogeneous DNA machine-triggered in situ formation of silver nanoclusters
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使用均质 DNA 机器触发原位形成银纳米簇检测 HIV/HCV 病毒 DNA
DOI：
10.1016/j.snb.2021.131041
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发表时间：
2021-11
期刊：
影响因子：
--
通讯作者：
Jian-Hua Wang
中科院分区：
文献类型：
--
作者：
Na Wu;Hui-Chao Zhang;Xu-Hong Sun;Feng-Na Guo;Li-Xia Feng;Ting Yang;Jian-Hua Wang
目前，早期疾病筛查仍是提高HIV/HCV感染者生存率的最有效途径。然而，痕量分析仍然是早期疾病筛查的一大挑战。本文通过结合Exo iii辅助靶循环扩增（ERA）和滚圈扩增（RCA），制备了一种新型银纳米簇（agnc）作为荧光探针，用于超灵敏的HIV/HCV DNA检测。作为该传感器的重要关键元件，RCA的挂锁探针（PLP）经过精心设计，组成了一个富鸟嘌呤（G-rich）区域。靶DNA首先与发夹DNA （hp-DNA）茎段杂交，触发Exo III酶切，释放并循环靶DNA，生成大量ssDNA片段。生成的ssDNA片段作为连接探针，与PLP杂交触发RCA过程，产生大量周期性重复的富胞嘧啶（C-rich）单元。在nabh4和AgNO3存在的情况下，会产生大量的agnc，从而产生强烈的荧光信号。在最优条件下，构建了检测HIV DNA的超灵敏DNA机，检测限为1.4 fM，线性范围为10 fM ~ 100 pM。更令人着迷的是，这台DNA机器可以通过构建模拟与或逻辑电路来智能区分多种病毒DNA。
Currently, the early disease screening is still the most effective way to increase the survival rate of the HIV/HCV infected patients. However, the trace analysis remains a great challenge for early disease screening. Herein, a novel silver nanocluster (AgNCs) isin situgenerated and served as fluorescence probes for the ultrasensitive HIV/HCV DNA detection by combing Exo III-assisted target recycling amplification (ERA) with rolling circle amplification (RCA). As an important key element of this sensor, the padlock probes (PLP) of RCA are exquisitely designed to compose of a guanine-rich (G-rich) region. Target DNA firstly hybridizes with the stem segment of the hairpin DNA (hp-DNA) and triggered the Exo III digestion, which releases and recycles the target and generate numerous ssDNA fragments. The generated ssDNA fragments act as ligation probes, which hybridize with PLP to trigger the RCA process, generating numerous periodically repeated cytosine-rich (C-rich) units. In the presence of NaBH4and AgNO3, numerous AgNCs arein situgenerated, thereby producing a strong fluorescent signal. Under optimal conditions, an ultrasensitive DNA machine is constructed for the detection of HIV DNA with a detection limit of 1.4 fM and a linear range of 10 fM to 100 pM. More fascinatingly, this DNA machine could intelligently distinguish multiple virus DNAs by the construction of analog AND and OR logic circuits.
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