Lab in a Tube: Ultrasensitive Detection of MicroRNAs at the Single-Cell Level and in Breast Cancer Patients Using Quadratic Isothermal Amplification

作者：Ruixue Duan, Xiaolei Zuo, Shutao Wang, Xiyun Quan, Dongliang Chen, Zhifei Chen, Lei Jiang, Chunhai Fan, Fan Xia · 发表于：Journal of the American Chemical Society · 年份：2013 · DOI：10.1021/ja311313b · 被引用次数：360 · 研究领域：Advanced biosensing and bioanalysis techniques、MicroRNA in disease regulation、RNA Interference and Gene Delivery

Through rational design of a functional molecular probe with high sequence specificity that takes advantage of sensitive isothermal amplification with simple operation, we developed a one-pot hairpin-mediated quadratic enzymatic amplification strategy for microRNA (miRNA) detection. Our method exhibits ultrahigh sensitivity toward miR-21 with detection limits of 10 fM at 37 °C and 1 aM at 4 °C, which corresponds to nine strands of miR-21 in a 15 μL sample, and it is capable of distinguishing among miRNA family members. More importantly, the proposed approach is also sensitive and selective when applied to crude extractions from MCF-7 and PC3 cell lines and even patient tissues from intraductal carcinoma and invasive ductal carcinoma of the breast.