Rolling circle amplification (RCA) -based biosensor system for the fluorescent detection of miR-129-2-3p miRNA

基于滚环扩增(RCA)的生物传感器系统用于荧光检测miR-129-2-3p miRNA

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作者:Yan Ye,Yao Lin,Zilin Chi,Jiasheng Zhang,Fan Cai,Youzhi Zhu,Dianping Tang,Qingqiang Lin

Abstract

Herein, a versatile fluorescent bioanalysis platform for sensitive and specific screening of target miRNA (miR-129-2-3p) was innovatively designed by applying target-induced rolling circle amplification (RCA) for efficient signal amplification. Specifically, miR-129-2-3p was used as a ligation template to facilitate its ligation with padlock probes, followed by an RCA reaction in the presence of phi29 DNA polymerase. The dsDNA fragments and products were stained by SYBR Green I and then detected by fluorescence spectrophotometry. As a result, miR-129-2-3p concentrations as low as 50 nM could be detected. Furthermore, the expression of miR-129-2-3p in breast cancer patients was about twice that in healthy people. Therefore, the results indicated that the RCA-based biosensor system could be a valuable platform for miRNA detection in clinical diagnosis and biomedical study.

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