Enzyme-Free Fluorescent Detection of MicroRNA in Clinical Samples by Catalytic Hairpin Assembly Coupled with Magnetic Bead-Confined 3D DNA Walking

IF 3.4 Q2 CHEMISTRY, ANALYTICAL
Jingyuan Yu, Lijuan Qi, Songchen Zhao, Dr. Xiaojun Zhang, Xudong Shang, Prof. Xintong Hu, Liguo Chen, Duo Wang, Prof. Yanfang Jiang, Prof. Yan Du
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Abstract

Catalytic hairpin assembly (CHA), as an enzyme-free isotheral nucleic acid amplification method, can easily cooperate with other amplification procedures to improve the sensitivity and accuracy of detection. Herein, we constructed a cascaded CHA sensing platform for breast cancer biomarker detection. Introducing a short double nucleic acid stand avoids the product of CHA1 to directly trigger the CHA2 reaction, which simplifies the design of the CHA hairpins. Compared with the single CHA2 reaction, the cascaded CHA biosensor activated by microRNA-155 holds nearly 10 times the amplification efficiency with detection limit down to 47.4 pM and quantifies the target in the range from 50 pM to 200 nM. Besides, the magnetic bead-confined CHA2 taking 3D DNA walking as the display form contributes to decreasing the environmental interference. As expected, the strategy sensitively distinguishes expression levels of microRNA-155 in different cell lines and cancer patients, which are consistent with the results of traditional qRT-PCR method. More importantly, simply adjusting the microRNA recognition sequence of CHA1 can extend the cascaded CHA platform to a wider detection range. Therefore, the robustness and efficiency of the approach enable the potential applications for detection of microRNA and early clinical disease diagnosis.

Abstract Image

催化发夹组装耦合磁珠约束三维DNA行走的临床样品中MicroRNA的无酶荧光检测
催化发夹组装法(Catalytic hairpin assembly, CHA)作为一种无酶的等温核酸扩增方法,可以方便地与其他扩增程序配合,提高检测的灵敏度和准确性。在此,我们构建了一个用于乳腺癌生物标志物检测的级联CHA传感平台。引入短双核酸支架,避免了CHA1的产物直接触发CHA2反应,简化了CHA发夹的设计。与单个CHA2反应相比,microRNA-155激活的级联CHA生物传感器扩增效率提高近10倍,检测限降至47.4 pM,可在50 pM ~ 200 nM范围内对目标进行定量。此外,以3D DNA行走为显示形式的磁珠约束CHA2有助于减少环境干扰。正如预期的那样,该策略灵敏地区分了microRNA-155在不同细胞系和癌症患者中的表达水平,这与传统的qRT-PCR方法的结果一致。更重要的是,只需调整CHA1的microRNA识别序列,就可以将级联CHA平台扩展到更广泛的检测范围。因此,该方法的鲁棒性和高效性使其在microRNA检测和早期临床疾病诊断方面具有潜在的应用前景。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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CiteScore
2.60
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