DNA nanoflowers encapsulating horseradish peroxidase as a signal amplification tag for rapid diagnosis in cytology specimens

作者全名：Xiang, Jiahui; Feng, Kai; Wan, Tao; He, Shiyin; Deng, Hongli; Li, Dairong

作者地址：[Xiang, Jiahui; Feng, Kai; Wan, Tao; He, Shiyin; Deng, Hongli; Li, Dairong] Chongqing Med Univ, Affiliated Hosp 1, Dept Resp & Crit Care Med, Chongqing 400016, Peoples R China; [Li, Dairong] 1 Youyi Rd, Chongqing 400016, Peoples R China

通信作者：Li, DR (通讯作者)，1 Youyi Rd, Chongqing 400016, Peoples R China.

来源：MICROCHEMICAL JOURNAL

ESI学科分类：CHEMISTRY

WOS号：WOS:001206985200001

JCR分区：Q1

影响因子：4.9

年份：2024

卷号：200

期号：　

开始页：　

结束页：　

文献类型：Article

关键词：Aptamer; One-step strategy; Immunocytochemistry; DNA nanoflowers; Cytological diagnosis

摘要：Rapid analysis of cytology samples obtained by minimally invasive sampling is essential in clinical diagnosis, yet existing diagnostic methods are often cumbersome and time-consuming. To rapidly detect the scanty number of cells in specimens, a multifunctional detection probe based on DNA nanoflower structure was prepared in this study, which possesses both signal output capability of fluorescence and oxidation reaction (colorimetric assay) for ultra -sensitive cancer cell detection. Firstly, DNA nanoflowers encapsulating horseradish peroxidase (HRPDNA Nanoflowers, HDFs) were prepared by the rolling circle amplification (RCA) reaction and then hybridized with the aptamers labeled with fluorophores to construct the multivalent aptamer HDFs (Multi-Aptamer-HDFs) with fluorescence effect and efficient cancer cell targeting. The Multi-Aptamer-HDFs probe can be used to detect fresh clinical cytology specimens directly for fluorescence observation with sensitivity and specificity of 88.64%, and 84.85%, respectively. In addition, Multi-Aptamer-HDFs has peroxidase activity due to the encapsulation of HRP, which can catalyze the oxidation of substrates such as ABTS and DAB for color development. Therefore, we applied Multi-Aptamer-HDFs to one-step staining of clinical cytology cell blocks with a sensitivity of 88.41% and a specificity of 83.33%, which required fewer steps and reagents with a significantly shorter time required for immunocytochemical staining. The multifunctional probe, Multi-Aptamer-HDFs, prepared in this study is expected to be better applied in the rapid cellular analyses and one-step immunostaining, which is expected to provide a new means for rapid on -site assessment and on -site preliminary diagnosis.

基金机构：Program of Chongqing Special Social Livelihood of Science and Technology Innovation [cstc2020 jscxmsxmX0063]; Graduate Scientific Research and Innovation Project of Chongqing [CYS22337]

基金资助正文：<BOLD>Funding</BOLD> This work was supported by the Program of Chongqing Special Social Livelihood of Science and Technology Innovation (NO. cstc2020 jscxmsxmX0063) , and the Graduate Scientific Research and Innovation Project of Chongqing (NO. CYS22337) .