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  • Title: Label-free detection of microRNA based on coupling multiple isothermal amplification techniques.
    Author: Zheng X, Niu L, Wei D, Li X, Zhang S.
    Journal: Sci Rep; 2016 Oct 25; 6():35982. PubMed ID: 27777399.
    Abstract:
    MicroRNA (miRNA) was a promising class of cancer biomarkers. Here we developed a label-free method for sensitive measurement of let-7d miRNA based on multiple amplification techniques. The primer will bind to the duplex strand DNA that was formed by stem-loop template and target let-7d to initiate strand displacement amplification (SDA) in tandem. The released single strand DNA will be a primer to bind the circular template to initiate rolling circle amplification (RCA). The products based on multiple amplifications will be detected by a standard fluorimeter with N-methyl mesoporphyrin IX (NMM) as the fluorescent indicator. The proposed method exhibited excellent selectivity and high sensitivity with a detection limit of as low as 1.5 × 10-13 M. Moreover, this methodology was used for the determination of biomolecules in real serum samples with satisfying results.
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