State Key Laboratory of Natural Medicines, Jiangsu Key Laboratory of TCM Evaluation and Translational Research, Research Center for Traceability and Standardization of TCMs, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing, 211198, PR China.
State Key Laboratory of Natural Medicines, Jiangsu Key Laboratory of TCM Evaluation and Translational Research, Research Center for Traceability and Standardization of TCMs, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing, 211198, PR China.
Anal Chim Acta. 2020 May 22;1112:72-79. doi: 10.1016/j.aca.2020.03.051. Epub 2020 Mar 27.
The upregulation or downregulation of microRNA-21 (miRNA-21) is closely related with drug-induced kidney injury (DIKI). As a potential and significant biomarker, the point-of-care testing (POCT) of miRNA-21 is worthy of attention and can provide essential information for clinical diagnosis. Hence, we design a portable and sensitive POCT assay for miRNA-21 using personal glucose meters (PGM). The whole operational system is constructed on streptavidin-coated magnetic beads (MBs) modified with substrate strands linked invertase and DNAzyme molecules each silenced by a locking strand. In the presence of miRNA-21, the locking strand can hybridize to miRNA-21, which originates the activation of the DNAzyme. The DNAzyme cleaves the substrate strands and induces the release of invertase from the surface of MBs. The separated invertase hydrolyzes sucrose to glucose which can be measured by PGM. The dual enzyme mediated catalyzation by DNAzyme and invertase therefore triggers the signal amplification. We establish a linear relationship between PGM and different concentration of miRNA-21 in the range of 100 fM to 1 pM. The limit of detection is 68.08 fM, which is comparable with some of the previous reports. The biosensor also exhibits excellent sequence selectivity, well-presented reproducibility and stability. Notably, by detecting miRNA-21 in urine, this method has been successfully used to predict DIKI and evaluate the protection effect of drugs on DIKI. Therefore, a dependable and low-cost POCT strategy for the detection of miRNA-21 is established, which is promising to supply valuable information for drug screening and evaluation of DIKI.
miRNA-21 的上调或下调与药物诱导的肾损伤（DIKI）密切相关。miRNA-21 作为一种有潜力和重要的生物标志物，其即时检测（POCT）值得关注，可以为临床诊断提供重要信息。因此，我们使用个人血糖仪（PGM）设计了一种用于 miRNA-21 的便携式和灵敏的 POCT 检测方法。整个操作系统构建在链霉亲和素包被的磁珠（MBs）上，修饰有与连接的蔗糖酶和 DNA 酶分子的底物链，每个分子都被锁定链沉默。在存在 miRNA-21 的情况下，锁定链可以与 miRNA-21 杂交，从而激活 DNA 酶。DNA 酶切割底物链，并诱导蔗糖酶从 MBs 表面释放。分离的蔗糖酶将蔗糖水解为葡萄糖，葡萄糖可以通过 PGM 进行测量。DNA 酶和蔗糖酶的双酶介导催化因此引发信号放大。我们在 100 fM 至 1 pM 的范围内建立了 PGM 与不同浓度 miRNA-21 之间的线性关系。检测限为 68.08 fM，与一些先前的报告相当。该生物传感器还表现出优异的序列选择性、良好的重现性和稳定性。值得注意的是，通过检测尿液中的 miRNA-21，该方法已成功用于预测 DIKI 并评估药物对 DIKI 的保护作用。因此，建立了一种可靠且低成本的 miRNA-21 检测 POCT 策略，有望为药物筛选和 DIKI 评估提供有价值的信息。
