化学
多重位移放大
核酸
环介导等温扩增
小RNA
分子生物学
聚合酶链反应
试剂
聚合酶
重组酶聚合酶扩增
DNA
肺癌
癌症
检出限
色谱法
生物化学
基因
生物
DNA提取
病理
遗传学
医学
物理化学
作者
Jingyi Chen,Jingzi Zhang,Qihui Xie,Chu Zhang,Yong Lü,Fan Zhang,Qingjiang Wang
标识
DOI:10.1016/j.aca.2024.342469
摘要
More and more studies have found that microRNAs (miRNAs) are markers of cancer, and detection of miRNAs is beneficial for early diagnosis and prognosis of cancer. In this paper, the isothermal strand displacement polymerase reaction (ISDPR), which is an enzyme-assisted nucleic acid amplification method, was studied to combine with microchip electrophoresis (MCE) for a simultaneously detection of two cancer related miRNAs named microRNA-21 (miR-21) and microRNA-221 (miR-221). In the ISDPR amplification, two different DNA hairpins (HPs) were specifically designed, so that miR-21 and miR-221 could respectively bind to HPs and started ISDPR amplification to generate two different products which were ultimately detected by MCE. The optimal conditions of ISDPR were carefully investigated, and the limits of detection (LOD) of miR-21 and miR-221 were as low as 0.35 fM and 0.25 fM (S/N = 3) respectively under these conditions. The human lung tumor cells and serum samples were analyzed by this ISDPR-MCE method and satisfactory results were obtained, which means that this method is of high sensitivity, high efficiency, low reagent consumption and simple operation in miRNAs detection.
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