State Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources, Guangxi Normal University, Guilin, 541004, China.
State Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources, Guangxi Normal University, Guilin, 541004, China; Key Laboratory of Ecology of Rare and Endangered Species and Environmental Protection, Ministry of Education, Guangxi Normal University, Guilin, 541004, China.
Talanta. 2020 Mar 1;209:120505. doi: 10.1016/j.talanta.2019.120505. Epub 2019 Oct 26.
Rapid and accurate detection of microRNA content in cells is of great significance. Here, an ultrasensitive microchip electrophoresis (MCE) method based on cascade chemiluminescence (CL) signal amplification was developed for the detection of microRNA-21 in cells. In this method, horseradish peroxidase labeled DNA was used as a signal probe, which could induce CL signal by the reaction of luminol and HO. Combining with two cyclic enzyme digestion reactions by T7 exonuclease, a large number of signal probes were degraded. By using MCE-CL as a separation and detection platform, an amplified CL signal peak was achieved. The developed MCE-CL method can detect miR-21 at a concentration as low as 1.0 × 10 M, which was enhanced by six orders of magnitude compared with those of conventional MCE-CL assay. This method has been applied for the detection of microRNA-21 in cell lysate, which show that there were significant differences of miR-21 among different types of cells, and the content in cancer cells was much higher than that in normal cells, which can be used for the identification of cancer cells. Therefore, the proposed method held great application potential in early diagnosis of tumor and biomedical research.
快速准确地检测细胞中的 microRNA 含量具有重要意义。在这里，开发了一种基于级联化学发光（CL）信号放大的超灵敏微芯片电泳（MCE）方法，用于检测细胞中的 microRNA-21。在该方法中，辣根过氧化物酶标记的 DNA 被用作信号探针，通过发光氨和 HO 的反应可以诱导 CL 信号。结合 T7 外切酶的两种循环酶消化反应，大量的信号探针被降解。通过使用 MCE-CL 作为分离和检测平台，实现了放大的 CL 信号峰。所开发的 MCE-CL 方法可以检测到浓度低至 1.0×10⁻¹⁰ M 的 miR-21，与传统的 MCE-CL 测定相比，其增强了六个数量级。该方法已应用于细胞裂解物中 microRNA-21 的检测，结果表明不同类型细胞中 miR-21 的含量存在显著差异，癌细胞中的含量明显高于正常细胞，可用于癌细胞的鉴定。因此，该方法在肿瘤的早期诊断和生物医学研究中具有很大的应用潜力。
