均相 SERS 生物传感器催化传感机制的设计、合理化和自动化,ACS Sensors

当前位置： X-MOL 学术 › ACS Sens. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

均相 SERS 生物传感器催化传感机制的设计、合理化和自动化
ACS Sensors ( IF 8.2 ) Pub Date : 2023-04-20 , DOI: 10.1021/acssensors.3c00175
Steven M Quarin ₁ , Amanda C Macke ₁ , Lyndsay N Kissell ₁ , Maria S Kelly ₁ , Ashan Dayananda ₁ , Joseph Ungvary ₁ , George Stan ₁ , Ruxandra I Dima ₁ , Pietro Strobbia ₁

Affiliation

当前的大流行表明我们需要灵敏且可部署的诊断技术。表面增强拉曼散射 (SERS) 传感器可以成为开发此类高级按需点 (PON) 诊断测试的理想解决方案。均质（无试剂）SERS 传感器通过直接响应目标而无需任何处理步骤来工作，使它们能够进行简单的一锅式检测，但它们的局限性在于可实现的灵敏度，与检测病毒生物标记所需的灵敏度相比是不够的。最近已将非共价 DNA 催化机制用于 SERS 测定中的催化扩增。这些进展使用催化发夹组装 (CHA) 和其他 DNA 自组装过程来开发具有更高灵敏度的传感机制。然而，这些机制尚未用于 OFF-to-ON 均质传感器，并且它们通常针对相同的生物标志物，这可能是由于机制设计的复杂性。仍然强烈需要具有均相机制和催化传感机制合理化的催化 SERS 传感器，以将这种传感策略转化为不同的目标和应用。我们开发并研究了一种均相 SERS 传感机制，该机制使用基于 DNA 自组装的催化放大。我们系统地研究了驱动催化机制的燃料链中三个结构域（内环、茎和立足点）的作用。在我们的研究中确定的热力学参数被用于构建一种算法，用于自动设计催化传感器，我们在与疟疾和 SARS-CoV-2 毒株相关的目标序列上验证了该算法。通过我们的机制，我们能够实现常规 DNA 的 20 倍扩增水平和使用锁定核酸 (LNA) 的 36 倍扩增水平，并在传感器检测限 (LOD) 中观察到相应的改进。我们还展示了针对与 omicron 变体相关的序列的传感器的单碱基序列特异性，针对 delta 变体目标进行了测试。这项关于均质 SERS 传感器催化放大的工作有可能通过改善 LOD 同时保持传感器的均质特性，使这种传感模式能够用于新的应用，例如传染病监测。我们还展示了针对与 omicron 变体相关的序列的传感器的单碱基序列特异性，针对 delta 变体目标进行了测试。这项关于均质 SERS 传感器催化放大的工作有可能通过改善 LOD 同时保持传感器的均质特性，使这种传感模式能够用于新的应用，例如传染病监测。我们还展示了针对与 omicron 变体相关的序列的传感器的单碱基序列特异性，针对 delta 变体目标进行了测试。这项关于均质 SERS 传感器催化放大的工作有可能通过改善 LOD 同时保持传感器的均质特性，使这种传感模式能够用于新的应用，例如传染病监测。

"点击查看英文标题和摘要"

Design, Rationalization, and Automation of a Catalytic Sensing Mechanism for Homogeneous SERS Biosensors

The current pandemic has shown that we need sensitive and deployable diagnostic technologies. Surface-enhanced Raman scattering (SERS) sensors can be an ideal solution for developing such advanced point-of-need (PON) diagnostic tests. Homogeneous (reagentless) SERS sensors work by directly responding to the target without any processing step, making them capable for simple one-pot assays, but their limitation is the achievable sensitivity, insufficient compared to what is needed for sensing of viral biomarkers. Noncovalent DNA catalysis mechanisms have been recently exploited for catalytic amplification in SERS assays. These advances used catalytic hairpin assembly (CHA) and other DNA self-assembly processes to develop sensing mechanisms with improved sensitivities. However, these mechanisms have not been used in OFF-to-ON homogeneous sensors, and they often target the same biomarker, likely due to the complexity of the mechanism design. There is still a strong need for a catalytic SERS sensor with a homogeneous mechanism and a rationalization of the catalytic sensing mechanism to translate this sensing strategy to different targets and applications. We developed and investigated a homogeneous SERS sensing mechanism that uses catalytic amplification based on DNA self-assembly. We systematically investigated the role of three domains in the fuel strand (internal loop, stem, and toehold), which drives the catalytic mechanism. The thermodynamic parameters determined in our studies were used to build an algorithm for automated design of catalytic sensors that we validated on target sequences associated with malaria and SARS-CoV-2 strains. With our mechanism, we were able to achieve an amplification level of 20-fold for conventional DNA and of 36-fold using locked nucleic acids (LNAs), with corresponding improvements observed in the sensor limit of detection (LOD). We also show a single-base sequence specificity for a sensor targeting a sequence associated with the omicron variant, tested against a delta variant target. This work on catalytic amplification of homogeneous SERS sensors has the potential to enable the use of this sensing modality in new applications, such as infectious disease surveillance, by improving the LOD while conserving the sensor’s homogeneous character.

更新日期：2023-04-20

点击分享查看原文

点击收藏

阅读更多本刊新发论文本刊介绍/投稿指南