Amplified single base-pair mismatch detection via aggregation of exonuclease-sheared gold nanoparticles.
Amplified single base-pair mismatch detection via aggregation of exonuclease-sheared gold nanoparticles.
复制标题
通过核酸外切酶剪切的金纳米颗粒聚集进行放大单碱基对错配检测
DOI：
10.1021/ac4040373
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发表时间：
2014-04-01
影响因子：
7.4
通讯作者：
Xiao, Yi
中科院分区：
文献类型：
--
作者：
Wu, Shuo;Liang, Pingping;Yu, Haixiang;Xu, Xiaowen;Liu, Yuan;Lou, Xinhui;Xiao, Yi
单核苷酸多态性（SNP）检测对于早期诊断、临床诊断和疾病预防是重要的，并且快速且灵敏的低成本SNP检测测定对于资源有限的临床环境是有价值的。我们提出了一个简单的平台，使敏感的，裸眼检测的SNP与最小的试剂和设备的要求，在室温下在15分钟内。SNP检测是在一个单一的管与一套DNA探针修饰的金纳米粒子（AuNPs），一个单一的核酸外切酶（Exo III），和目标的问题。Exo III的脱嘌呤核酸内切酶活性差异化地处理AuNP结合的探针和完全匹配或含有单碱基错配的DNA靶之间的杂交双链体。对于完全匹配的靶标，Exo III的核酸外切酶活性促进了靶标再循环的过程，该过程快速地从颗粒剪切DNA探针，产生AuNP聚集诱导的颜色变化，而对于错配的靶标则不会发生这种变化。这种颜色变化很容易用低至2 nM的靶观察到，比在充分优化的反应条件下用未修饰的AuNP进行可靠的肉眼观察所需的靶浓度低100倍。我们进一步证明，该系统可以有效地区分一系列不同的错配。
Single nucleotide polymorphism (SNP) detection is important for early diagnosis, clinical prognostics, and disease prevention, and a rapid and sensitive low-cost SNP detection assay would be valuable for resource-limited clinical settings. We present a simple platform that enables sensitive, naked-eye detection of SNPs with minimal reagent and equipment requirements at room temperature within 15 min. SNP detection is performed in a single tube with one set of DNA probe-modified gold nanoparticles (AuNPs), a single exonuclease (Exo III), and the target in question. Exo III’s apurinic endonucleolytic activity differentially processes hybrid duplexes between the AuNP-bound probe and DNA targets that are perfectly matched or contain a single-base mismatch. For perfectly matched targets, Exo III’s exonuclease activity facilitates a process of target recycling that rapidly shears DNA probes from the particles, generating an AuNP aggregation-induced color change, whereas no such change occurs for mismatched targets. This color change is easily observed with as little as 2 nM of target, 100-fold lower than the target concentration required for reliable naked eye observation with unmodified AuNPs in well-optimized reaction conditions. We further demonstrate that this system can effectively discriminate a range of different mismatches.
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影响因子：
14.9
作者：
Bellamy SR;Kovacheva YS;Zulkipli IH;Halford SE
通讯作者：
Halford SE
DOI：
10.1089/109065700316408
发表时间：
2000-03-01
期刊：
GENETIC TESTING
影响因子：
--
作者：
Kurg, A;Tonisson, N;Metspalu, A
通讯作者：
Metspalu, A
影响因子：
15
作者：
Shen, Wei;Deng, Huimin;Gao, Zhiqiang
通讯作者：
Gao, Zhiqiang
影响因子：
46.9
作者：
Dubertret, B;Calame, M;Libchaber, AJ
通讯作者：
Libchaber, AJ
影响因子：
7
作者：
Heid, CA;Stevens, J;Williams, PM
通讯作者：
Williams, PM