Han Yu, Li Jialin, Li Man, An Ran, Zhang Xu, Cai Sheng
College of Pharmacy, Jilin Medical University, Jilin 132013, China.
Institute of Drug Metabolism and Pharmaceutical Analysis, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research, Zhejiang University, Hangzhou 310058, China.
Molecules. 2024 Dec 6;29(23):5782. doi: 10.3390/molecules29235782.
The association between microRNA (miRNA) and various diseases has been established; miRNAs have the potential to be biomarkers for these diseases. Nevertheless, the challenge of correctly quantifying an miRNA arises from its low abundance and a high degree of family homology. Therefore, in the present study, we devised a chemiluminescence (CL) detection method for miRNAs, known as the hybridization chain reaction (HCR)-CL, utilizing the enzyme-free signal amplification technology of HCR. The proposed methodology obviates the need for temperature conversion and offers a straightforward procedure owing to the absence of enzymatic participation, and the lumino-HO-mediated CL reaction occurs at a high rate. The technique successfully detected 2.5 amol of the target analyte and 50 amol of miR-146b in a 1% concentration of human serum. In summary, the method developed in this study is characterized by its ease of operation, cost-effectiveness, remarkable analytical prowess, and ability to detect miRNA without the need for total RNA extraction from serum samples. This method is expected to be widely used for biological sample testing in clinical settings.
微小RNA（miRNA）与多种疾病之间的关联已被确立；miRNA有潜力成为这些疾病的生物标志物。然而，由于miRNA丰度低且家族同源性高，准确量化miRNA面临挑战。因此，在本研究中，我们利用杂交链式反应（HCR）的无酶信号放大技术，设计了一种用于miRNA的化学发光（CL）检测方法，即HCR-CL。所提出的方法无需温度转换，且由于没有酶的参与，操作流程简单，并且由鲁米诺-HO介导的CL反应速率很高。该技术在1%浓度的人血清中成功检测到2.5 amol的目标分析物和50 amol的miR-146b。总之，本研究开发的方法具有操作简便、成本效益高、出色的分析能力以及无需从血清样本中提取总RNA就能检测miRNA的特点。预计该方法将在临床环境中广泛用于生物样本检测。
