Ligation-promoted hyperbranched rolling circle amplification enables ultrasensitive detection of microRNA in clinical specimens
Ligation-promoted hyperbranched rolling circle amplification enables ultrasensitive detection of microRNA in clinical specimens
复制标题
连接促进的超支化滚环扩增能够超灵敏地检测临床样本中的 microRNA
DOI：
10.1016/j.snb.2018.09.058
复制
发表时间：
2018-12-20
影响因子：
8.4
通讯作者：
Zhou, Xiang
中科院分区：
文献类型：
--
作者：
Zhang, Xiaoe;Liu, Yinong;Zhou, Xiang
据报道，微小RNA（miRNA）是多种疾病和癌症中重要的生物标志物候选物和治疗靶点。然而，从临床样本中检测miRNA的超灵敏、简便、等温、低成本且可靠的方法仍然是挑战。在此，我们基于一种新型连接促进的超支化滚环扩增（L - HRCA）建立了一种等温、高灵敏度和特异性的miRNA分析方法。与T4连接酶辅助的靶标 - 引物滚环扩增（BRCA）相比，该策略的灵敏度提高了10³倍。L - HRCA的优异特异性能够区分目标miRNA和阴性miRNA。该方法被应用于分析肝细胞癌中miR - 21的表达水平。这些结果表明，L - HRCA策略将成为早期临床诊断中定量miRNA的一种有前景的工具。
MiRNA is reported as an important biomarker candidate and therapeutic target in a variety of diseases and cancers. However, ultrasensitive, straightforward, isothermal, low-cost and credible methods for the detection of miRNA from clinical specimens still remain challenges. Here, we establish an isothermal, highly sensitive and specific method for miRNA analysis based on a novel ligation-promoted hyperbranched rolling circle amplification (L-HRCA). This strategy exhibits 10(3)-fold sensitivity enhancement compared to T4 ligase-assisted targetprimer BRCA. The excellent specificity of L-HRCA allows for discriminating target miRNA from negative miRNAs. This method is applied to analyse the expression level of miR-21 in hepatocellular carcinoma. These results indicated that the L-HRCA strategy will become a promising tool to quantify miRNA in early clinical diagnostics.