一种通过复制杂交链反应的可相互放大的荧光传感平台，用于承载连接的多 Ag 纳米团簇作为信号报告器,Analytical Chemistry

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一种通过复制杂交链反应的可相互放大的荧光传感平台，用于承载连接的多 Ag 纳米团簇作为信号报告器
Analytical Chemistry ( IF 6.7 ) Pub Date : 2022-11-15 , DOI: 10.1021/acs.analchem.2c03782
Mengdie Li ₁ , Jiayang He ₁ , Xin Shang ₁ , Chunli Yang ₁ , Yuqing Zhang ₁ , Siyu Zuo ₁ , Ruo Yuan ₁ , Wenju Xu ₁

Affiliation

探索用于相互扩增的杂交链式反应 HCR ( r HCR) 的复制在生物传感和生物分析中很有趣。在此，我们开发了一个基于r HCR 的荧光平台，该平台由特定 DNA 触发器 ( T ) 和T类似扩增子 ( T * ) 的组合操纵，从而连接多个绿色发射 Ag 纳米团簇 ( mg AgNCs) 用于可放大信号读出。执行具有顺序补码的四个精心设计的发夹（H1 识别T 、H2、H3 和 H4）来操作r HCR。H1/H3 的末端合并以杂交抑制链 ( I). mg AgNCs的母体支架分为两个部分（C ₄ AC ₄ T 和 C ₃ GT ₄），分别悬垂在 H2/H4 中。T的存在通过四个反应性发夹的交叉杂交激活第一个 HCR 放大器以形成 HCR 双链体。复合体的下一次入侵 ( T * ·I ) 驱使I杂交 H1/H3 中的串联重复序列，使得置换的T*充当T以催化第二个放大器r HCR，以快速反应动力学反馈更多的发夹组件. 在共享rHCR 聚合物、H2/H4 中的母体支架 (C ₄ AC ₄ TC ₃ GT ₄ ) 共同串联在一起，以优先聚集mg AgNCs 加合物，它们协同发出巨大的T响应荧光信号。由于 HCR 产品中T的定位，引入了另一种扩增子T*以通过 DNA 链置换驱动r HCR 进程，从而产生更多的发射体成核位点。因此，非酶促r的合理组合HCR 和无标记荧光多联体会产生相互信号放大，实现低至飞摩尔浓度的简化、快速和高度灵敏的检测。

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A Reciprocal-Amplifiable Fluorescence Sensing Platform via Replicated Hybridization Chain Reaction for Hosting Concatenated Multi-Ag Nanoclusters as Signal Reporter

Exploring the replication of hybridization chain reaction HCR (rHCR) for reciprocal amplification is intriguing in biosensing and bioanalysis. Herein, we develop a rHCR-based fluorescence platform that is manipulated by the combination of a specific DNA trigger (T) and a T-analogous amplicon (T*), thereby concatenating multi green-emissive Ag nanoclusters (mgAgNCs) for amplifiable signal readout. Four well-designed hairpins (H1 recognizing T, H2, H3, and H4) with sequential complements are executed to operate rHCR. The termini of H1/H3 are merged to hybridize an inhibiting strand (I). The parent scaffold for mgAgNCs is separated into two splits (C₄AC₄T and C₃GT₄) that are individually overhung in H2/H4. The presence of T activates the first HCR amplifier through cross-hybridization of four reactive hairpins for forming HCR duplexes. The next invasion of a complex (T*·I) drives I to hybridize the tandem repeats in H1/H3, so that the displaced T* functions as T to catalyze the second amplifier rHCR for feeding back more hairpin assemblies with rapid reaction kinetics. In the shared rHCR polymers, the parent scaffolds (C₄AC₄TC₃GT₄) in H2/H4 are collectively concatenated for the preferential clustering of mgAgNCs adducts, which cooperatively emit enormous T-responsive fluorescence signal. Because of the localization of T in HCR products, an alternative amplicon T* is introduced to drive rHCR progress via DNA strand displacement, generating more nucleating sites of emitters. Thus, the rational combination of nonenzymatic rHCR and label-free fluorescent concatemers would create a reciprocal signal amplification, achieving a simplified, rapid, and highly sensitive assay down to femtomolar concentrations.

更新日期：2022-11-15

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