Dumbbell structure probe-triggered rolling circle amplification (RCA)-based detection scaffold for sensitive and specific neonatal infection-related small extracellular vesicle (sEV) detection

作者全名："Yang, Zeping; She, Dong; Sun, Chunhong; Gong, Mingwei; Rong, Yuan"

作者地址："[Yang, Zeping; She, Dong; Sun, Chunhong; Gong, Mingwei; Rong, Yuan] Chongqing Med Univ, Dept Crit Care Med ICU, Affiliated Hosp 3, Chongqing 401120, Peoples R China"

通信作者："Rong, Y (通讯作者)，Chongqing Med Univ, Dept Crit Care Med ICU, Affiliated Hosp 3, Chongqing 401120, Peoples R China."

来源：ANALYTICAL METHODS

ESI学科分类：CHEMISTRY

WOS号：WOS:000776765900001

JCR分区：Q2

影响因子：3.1

年份：2022

卷号：14

期号：15

开始页：1534

结束页：1539

文献类型：Article

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摘要："Small extracellular vesicles (sEVs) have been reported to play important roles in cell-to-cell communication and are promising biomarkers for the early diagnosis of infections. Therefore, it is in high demand to develop a method that can integrate easy-to-operate sEV isolation and sensitive quantification. We herein propose a novel detection scaffold for sEV isolation via low-speed centrifugation and the quantification of sEVs through DNAzyme-based signal amplification. The detection scaffold is established through dumbbell probe-based RCA (rolling circle amplification), containing repeated CD63 aptamer sections and DNAzyme sections. The original state of the DNAzyme section is locked in a hairpin structure in the detection scaffold. In the presence of sEVs, the CD63 aptamer recognizes and binds with sEVs, leading to the aggregation of sEVs, which can be isolated by low-speed centrifugation and the exposure of the DNAzyme section. After the catalytic fluorescence signal generation from the DNAzyme-based molecular beacon (MB) cleavage, the method exhibited a detection range of 10(2) to 10(6) particles per mu L. Considering the high sensitivity and wash-free and easy-to-operate features, the strategy reported herein paves a new avenue for the effective determination of sEVs and other membrane biomolecules in fundamental and applied research."

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