A Novel Design Combining Isothermal Exponential Amplification and Gold-Nanoparticles Visualization for Rapid Detection of miRNAs.
A Novel Design Combining Isothermal Exponential Amplification and Gold-Nanoparticles Visualization for Rapid Detection of miRNAs.
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结合等温指数扩增和金纳米粒子可视化的新颖设计，用于快速检测 miRNA
DOI：
10.3390/ijms19113374
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发表时间：
2018-10-28
影响因子：
5.6
通讯作者：
Guan Y
中科院分区：
文献类型：
--
作者：
Jiang J;Zhang B;Zhang C;Guan Y
微小RNA（miRNAs）在广泛的生物过程中发挥着重要作用，其异常表达与多种疾病相关。miRNAs的水平可作为细胞事件或疾病诊断的有用生物标志物；因此，对微小RNA进行灵敏且选择性的检测对于理解miRNAs的生物学功能、癌症的早期诊断以及发现药物新靶点具有重要意义。然而，传统的miRNAs检测方法通常费力且耗时，灵敏度较低。在此，我们基于等温指数扩增反应（EXPAR）和金纳米粒子（AuNP）标记的DNA探针相结合的方法，开发了一种简单、快速、超灵敏的比色检测法用于检测miRNAs（以let - 7a作为模型分析物）。在该检测法中，在等温条件下，let - 7a的存在通过EXPAR转化为报告分子Y。随后报告分子Y与AuNP标记的DNA探针进行夹心杂交，产生从红色到紫色的颜色变化。换句话说，如果报告分子Y与AuNP标记的DNA探针互补，DNA功能化的AuNPs将会聚集，导致溶液颜色从红色变为紫色/蓝色，而当AuNP标记的DNA探针与报告分子Y不匹配时，溶液保持红色。这种检测法代表了一种简单、省时的技术，由于比色变化，其结果可以用肉眼直接观察。该方法具有极高的灵敏度，在最佳条件下，检测限为4.176 aM，检测范围从1 nM到1 aM。该方法对于区分let - 7 miRNA家族成员之间甚至单核苷酸的差异也显示出高选择性。值得注意的是，它与迄今为止报道的最灵敏的方法相当，因此为直接检测let - 7a miRNA的标准方法提供了一种有前景的替代方法。重要的是，通过与特定模板结合，不同的miRNAs可以转化为相同的报告分子Y，它可以与同一组AuNP标记的DNA探针杂交形成夹心杂交体。在目标miRNA存在的情况下，可以观察到溶液的颜色变化。这种技术有可能成为评估miRNAs水平的常规方法，不仅适用于let - 7，也适用于癌症早期的各种miRNAs。此外，它在生物医学研究和临床诊断以及现场条件下的诊断或监测项目中也可能是一种有用的工具。
MicroRNAs (miRNAs) play important roles in a wide range of biological processes, and their aberrant expressions are associated with various diseases. The levels of miRNAs can be useful biomarkers for cellular events or disease diagnosis; thus, sensitive and selective detection of microRNAs is of great significance in understanding biological functions of miRNAs, early-phase diagnosis of cancers, and discovery of new targets for drugs. However, traditional approaches for the detection of miRNAs are usually laborious and time-consuming, with a low sensitivity. Here, we develop a simple, rapid, ultrasensitive colorimetric assay based on the combination of isothermal Exponential Amplification Reaction (EXPAR) and AuNP-labeled DNA probes for the detection of miRNAs (taking let-7a as a model analyte). In this assay, the presence of let-7a is converted to the reporter Y through EXPAR under isothermal conditions. The subsequent sandwich hybridization of the reporter Y with the AuNP-labeled DNA probes generates a red-to-purple color change. In other words, if the reporter Y is complementary to the AuNP-labeled DNA probes, the DNA-functionalized AuNPs will be aggregated, resulting in the change of solution color from red to purple/blue, while when the AuNP-labeled DNA probes are mismatched to the reporter Y, the solution remains red. This assay represents a simple, time-saving technique, and its results can be visually detected with the naked eye due to the colorimetric change. The method provides superior sensitivity, with a detection limit of 4.176 aM over a wide range from 1 nM to 1 aM under optimal conditions. The method also shows high selectivity for discriminating even single-nucleotide differences between let-7 miRNA family members. Notably, it is comparable to the most sensitive method reported to date, thus providing a promising alternative to standard approaches for the direct detection of let-7a miRNA. Importantly, through combination with specific templates, different miRNAs can be converted to the same reporter Y, which can hybridize with the same set of AuNP-labeled DNA probes to form sandwich hybrids. The color change of the solution can be observed in the presence of the target miRNA. This technique has potential as a routine method for assessing the levels of miRNAs, not only for let-7, but also for various miRNAs in the early phase of cancers. In addition, it can be a useful tool in biomedical research and clinical diagnosis, as well as diagnosis or surveillance programs in field conditions.
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影响因子：
14.9
作者：
Kiesling T;Cox K;Davidson EA;Dretchen K;Grater G;Hibbard S;Lasken RS;Leshin J;Skowronski E;Danielsen M
通讯作者：
Danielsen M
通过指数等温扩增超灵敏检测 microRNA
影响因子：
16.6
作者：
Jia, Hongxia;Li, Zhengping;Cheng, Yongqiang
通讯作者：
Cheng, Yongqiang
DOI：
10.1073/pnas.0730811100
发表时间：
2003-04-15
影响因子：
11.1
作者：
Van Ness, J;Van Ness, LK;Galas, DJ
通讯作者：
Galas, DJ
影响因子：
12.6
作者：
Wang, Ke;Zhang, Kai;Zhou, Fanfan
通讯作者：
Zhou, Fanfan
影响因子：
12.6
作者：
Yu, Yanyan;Chen, Zuanguang;Yao, Meicun
通讯作者：
Yao, Meicun