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Express and sensitive detection of multiple miRNAs via DNA cascade reactors functionalized photonic crystal array
Science China Chemistry ( IF 9.6 ) Pub Date : 2020-04-08 , DOI: 10.1007/s11426-020-9712-y
Yingfei Wang , Yuyi Li , Yue Zhang , Kewei Ren , Huangxian Ju , Ying Liu

Array based detection techniques with fluorescence signal reading is a powerful tool for multiple targets analysis. However, when applied fluorescence array for microRNA detection, time-consuming multi-steps surface signal amplification is usually required due to the low abundance of microRNA in total RNA expressions, which impairs detection efficiency and limits its application in point of care test (POCT) manner. Herein, DNA cascade reactors (DCRs) functionalized photonic crystal (PC) array was fabricated for express and sensitive detections of miRNA-21 and miRNA-155. DCRs were assembled by interval conjugation of self-quenched hairpin DNA probes to single strand DNA nanowire synthesized by rolling circle amplification, which generated cascade DNA hybridization reactions in response to target miRNA with instant fluorescence recovery signal. PC array patterns with multi-structure colors further amplified fluorescence with their respective photonic bandgaps (PBGs) matching with the emission peaks of fluorescence molecules labelled on DCRs. The as-prepared DCRs functionalized PC array demonstrated express and sensitive simultaneous detections of miRNA-21 and miRNA-155 with hundreds fM detection limits only in 15 min, and was successfully applied in fast quantifications of low abundance miRNAs from cell lysates and spiked miRNAs from human serum, which would hold great potential for disease diagnosis and therapeutic effect monitoring with a POCT manner.



中文翻译:

通过DNA级联反应器功能化的光子晶体阵列快速灵敏地检测多个miRNA

具有荧光信号读取功能的基于阵列的检测技术是进行多目标分析的强大工具。但是,当将荧光阵列应用于微RNA检测时,由于总RNA表达中微RNA的丰度低,通常需要耗时的多步表面信号放大,这会削弱检测效率并限制其在即时检测(POCT)中的应用方式。在此,制造了DNA级联反应器(DCR)功能化的光子晶体(PC)阵列,用于表达和灵敏检测miRNA-21和miRNA-155。通过将自淬灭的发夹DNA探针与通过滚环扩增合成的单链DNA纳米线间隔缀合来组装DCR,DCR响应于具有即时荧光恢复信号的靶标miRNA而产生级联DNA杂交反应。具有多色结构的PC阵列图案进一步放大了荧光,其各自的光子带隙(PBG)与DCR上标记的荧光分子的发射峰相匹配。所制备的DCRs功能化PC阵列仅在15分钟内即可表达和灵敏地同时检测出数百个fM检测限的miRNA-21和miRNA-155,并成功应用于细胞裂解液中低丰度miRNA和来自样品中的加标miRNA的快速定量人血清,以POCT方式在疾病诊断和治疗效果监测方面具有巨大潜力。

更新日期:2020-04-22
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