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

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• 作者：Ruixue Duan ; Xiaolei Zuo ; Shutao Wang ; Xiyun Quan ; Dongliang Chen ; Zhifei Chen ; Lei Jiang ; Chunhai Fan ; Fan Xia
• 刊名：The Journal of the American Chemical Society
• 出版年：2013
• 出版时间：March 27, 2013
• 年：2013
• 卷：135
• 期：12
• 页码：4604-4607
• 全文大小：290K
• 年卷期：v.135,no.12(March 27, 2013)
• ISSN：1520-5126

文摘

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 掳C and 1 aM at 4 掳C, which corresponds to nine strands of miR-21 in a 15 渭L 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.