Conventional PCR assisted single-component assembly of spherical nucleic acids for simple colorimetric detection of SARS-CoV-2.
Conventional PCR assisted single-component assembly of spherical nucleic acids for simple colorimetric detection of SARS-CoV-2.
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DOI：
10.1016/j.snb.2020.128971
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发表时间：
2021-02-01
期刊：
影响因子：
--
通讯作者：
Ezati R
中科院分区：
文献类型：
--
作者：
Karami A;Hasani M;Azizi Jalilian F;Ezati R
PCR的扩增能力与SNA组件的可编程性相结合，用于检测COVID-19。基于SARS-CoV-2 RNA的E基因的接头单组分组装系统提供了灵敏的比色响应。在PCR扩增过程中，回文接头基于5′-外切核酸酶活性被切割。阳性和阴性病毒COVID-19样本通过肉眼的不同颜色简单区分。持续识别疑似传染病例对于控制最近由新型人类冠状病毒SARS-CoV-2（严重急性呼吸综合征冠状病毒2）引起的大流行至关重要。由于缺乏资源和基础设施，实时聚合酶链反应（实时PCR）技术在一些社区无法容易地大规模实施。在这里，我们报告了一个简单的比色策略，来自基于连接基的单组分组装金纳米颗粒核心球形核酸（AuNP-core SNA）的PCR产物的SARS-CoV-2核糖核酸（RNA）模板的视觉检测。基于SARS-CoV-2特异性E基因设计了回文接头，以将相同的胶体SNA编程为大的组装体，沿着具有明显的红色到紫色的颜色变化。在常规PCR反应中，接头作为SARS-CoV-2 RNA的探针。在存在正确模板的情况下，与靶扩增子内的短区域互补的回文接头被脱氧核糖核酸（DNA）聚合酶的5′-外切核酸酶活性切割。在扩增过程中回文接头的切割抑制SNA的单组分组装形成。因此，阳性和阴性病毒样品在PCR后比色试验中分别产生简单的红色和紫色。从实验室确诊的SARS-CoV-2感染病例中获得的样品的评估表明，我们的检测方法在灵敏度上可以与实时PCR方法相媲美。
The amplification power of PCR is coupled with programmability of SNA assembly for detection of COVID-19. A linker-based single-component assembly system based on E gene of SARS-CoV-2 RNA provided sensitive colorimetric response. The palindromic linker is cleaved based on 5′-exonuclease activity in the course of PCR amplification. The positive and negative viral COVID-19 samples are simply distinguished via different colors by naked eyes. Continuous identification of suspected infectious cases is crucial to control the recent pandemic caused by the novel human coronavirus SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2). Real-time polymerase chain reaction (real-time PCR) technology cannot be implemented easily and in large scale in some communities due to lack of resources and infrastructures. Here, we report a simple colorimetric strategy derived from linker-based single-component assembly of gold nanoparticle-core spherical nucleic acids (AuNP-core SNAs) for visual detection of PCR products of SARS-CoV-2 ribonucleic acid (RNA) template. A palindromic linker is designed based on SARS-CoV-2 specific E gene to program the identical colloidal SNAs into large assemblies along with a distinct red-to-purple color change. The linker acts as a probe of SARS-CoV-2 RNA in conventional PCR reaction. In the presence of the correct template the palindromic linker, which is complementary to a short region within the target amplicon, is cleaved by 5′-exonuclease activity of deoxyribonucleic acid (DNA) polymerase. Cleavage of the palindromic linker during the amplification process inhibits the single-component assembly formation of SNAs. So, positive and negative viral samples produce simply red and purple colors in the post PCR colorimetric test, respectively. Evaluation of the samples obtained from cases with laboratory-confirmed SARS-CoV-2 infection revealed that our assay can rival with real-time PCR method in sensitivity.
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影响因子：
64.8
作者：
Mirkin, CA;Letsinger, RL;Storhoff, JJ
通讯作者：
Storhoff, JJ
DOI：
10.1073/pnas.1809667116
发表时间：
2019-02-12
影响因子：
11.1
作者：
Hulswit, Ruben J. G.;Lang, Yifei;de Groot, Raoul J.
通讯作者：
de Groot, Raoul J.
影响因子：
2.9
作者：
Lu, Le;Jia, Hui;Li, Jinming
通讯作者：
Li, Jinming
影响因子：
3
作者：
Dirks, RM;Pierce, NA
通讯作者：
Pierce, NA
影响因子：
10.2
作者：
Dirks, Robert M.;Bois, Justin S.;Pierce, Niles A.
通讯作者：
Pierce, Niles A.