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基于氧化石墨烯(GO)对荧光标记单链DNA探针的荧光猝灭效应以及双链特异性核酸酶(DSN)选择性切割DNA/RNA杂合结构中DNA单链的特性,本文建立了一种新型恒温信号放大方法用于microRNA(miRNA)的高灵敏度检测.靶标miRNA首先与荧光DNA探针杂交,DSN能够特异性地将杂合双链中的DNA探针水解为碎片但不会降解miRNA,GO对酶切产生的寡核苷酸碎片吸附能力显著降低,使得荧光基团远离GO表面而不被猝灭.释放出的miRNA可再次发生与荧光DNA探针杂交、DSN酶切等反应,如此反复,可实现恒温条件下一个miRNA分子与多个探针杂交、酶切、释放荧光基团的循环过程,最终体系的荧光信号得到显著放大,通过记录体系的荧光信号即可实现对靶标miRNA的灵敏检测.
Based on the fluorescence quenching effect of graphene oxide (GO) on fluorescently labeled single-stranded DNA probes and the characteristic of single-strand DNA cleavage in DNA / RNA hybrids by double-stranded specific nucleases (DSNs) A Novel Thermostatic Signal Amplification Method for Highly Sensitive Detection of MicroRNAs (miRNAs) Target miRNAs are first hybridized to fluorescent DNA probes that specifically hydrolyze DNA probes in hybrid duplexes into fragments without degrading them , GO on the enzymatic cleavage of oligonucleotide fragments significantly reduced adsorption capacity, making the fluorophores away from the GO surface without being quenched.The miRNA released again with fluorescent DNA probe hybridization, DSN digestion and other reactions, So repeatedly, under the condition of constant temperature, a miRNA molecule can be hybridized with a plurality of probes to digest and release the fluorescent group, and the fluorescence signal of the final system is significantly amplified. By recording the fluorescent signal of the system, the target miRNA Sensitive test.