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Point-of-Care Testing of MicroRNA based on Personal Glucose Meter and Dual Signal Amplification to Evaluate Drug-Induced Kidney Injury
Analytica Chimica Acta ( IF 5.7 ) Pub Date : 2020-05-01 , DOI: 10.1016/j.aca.2020.03.051
Xitong Huang 1 , Jiwei Li 1 , Mi Lu 1 , Wangning Zhang 1 , Zhiming Xu 1 , Bo-Yang Yu 1 , Jiangwei Tian 1
Affiliation  

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.

中文翻译:

基于个人血糖仪和双信号放大的 MicroRNA 即时检测以评估药物诱导的肾损伤

microRNA-21(miRNA-21)的上调或下调与药物性肾损伤(DIKI)密切相关。作为一种潜在且重要的生物标志物,miRNA-21的即时检测(POCT)值得关注,可为临床诊断提供重要信息。因此,我们使用个人血糖仪 (PGM) 为 miRNA-21 设计了一种便携式且灵敏的 POCT 检测方法。整个操作系统是在链霉亲和素包被的磁珠 (MB) 上构建的,这些磁珠是用底物链连接的转化酶和 DNAzyme 分子修饰的,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,从而引发 DNAzyme 的激活。DNAzyme 切割底物链并诱导转化酶从 MB 表面释放。分离的转化酶将蔗糖水解为葡萄糖,可通过 PGM 测量。因此,由 DNAzyme 和转化酶介导的双酶催化会触发信号放大。我们在 100 fM 到 1 pM 的范围内建立了 PGM 与不同浓度的 miRNA-21 之间的线性关系。检测限为 68.08 fM,与之前的一些报告相当。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 POCT 检测 miRNA-21 策略,有望为 DIKI 的药物筛选和评估提供有价值的信息。
更新日期:2020-05-01
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