Country Department of Clinical Laboratory Medicine, Shanghai Chest Hospital, Shanghai Jiao Tong University, Shanghai, 200030, China.
Shanghai Institute of Thoracic Oncology, Shanghai Chest Hospital, Shanghai Jiao Tong University, Shanghai, 200030, China.
Small. 2022 Jun;18(22):e2200784. doi: 10.1002/smll.202200784. Epub 2022 Mar 25.
Circulating microRNAs (miRNAs) can be used as noninvasive biomarkers and are also found circulating in body fluids such as blood. Dysregulated miRNA expression is associated with many diseases, including non-small cell lung cancer (NSCLC), and the miRNA assay is helpful in cancer diagnosis, prognosis, and monitoring. In this work, a versatile electrochemical biosensing system is developed for miRNA detection by DNAzyme-cleavage cycling amplification and hybridization chain reaction (HCR) amplification. With cleavage by Mn targeted DNAzyme, DNA-walker can move along the predesigned DNA tracks and contribute to the transduction and enhancement of signals. For the electrochemical process, the formation of multiple G-quadruplex-incorporated long double-stranded DNA (dsDNA/G-quadruplex) structures is triggered through HCR amplification. The introduction of G-quadruplex allows sensitive measurement of miRNA down to 5.68 fM with good specificity. Furthermore, by profiling miRNA in the NSCLC cohort, this designed strategy shows high efficiency (area under the curve (AUC) of 0.879 using receiver operating characteristic (ROC) analysis) with the sensitivity of 80.0% for NSCLC early diagnosis (stage I). For the discrimination of NSCLC and benign disease, the assay displays an AUC of 0.907, superior to six clinically-acceptable protein tumor markers. Therefore, this platform holds promise in clinical application toward NSCLC diagnosis and prognosis.
循环 microRNAs（miRNAs）可用作非侵入性生物标志物，也存在于血液等体液中循环。miRNA 表达失调与许多疾病有关，包括非小细胞肺癌（NSCLC），miRNA 检测有助于癌症的诊断、预后和监测。在这项工作中，开发了一种多功能电化学生物传感系统，用于通过 DNA 酶切割循环扩增和杂交链式反应（HCR）扩增检测 miRNA。通过 Mn 靶向 DNA 酶的切割，DNA 行走器可以沿着预设的 DNA 轨道移动，有助于信号的转导和增强。对于电化学过程，通过 HCR 扩增触发多个包含 G-四链体的长双链 DNA（dsDNA/G-四链体）结构的形成。引入 G-四链体可以通过灵敏测量 miRNA 来实现，其下限低至 5.68 fM，具有良好的特异性。此外，通过对 NSCLC 队列中的 miRNA 进行分析，该设计策略显示出高效率（使用接收者操作特征（ROC）分析的 AUC 为 0.879），对 NSCLC 早期诊断（I 期）的灵敏度为 80.0%。对于 NSCLC 和良性疾病的区分，该检测方法的 AUC 为 0.907，优于六种临床可接受的蛋白肿瘤标志物。因此，该平台有望在 NSCLC 诊断和预后的临床应用中发挥作用。
