Rolling circle amplification (RCA) -based biosensor system for the fluorescent detection of miR-129-2-3p miRNA
Yan Ye,
Yao Lin,
Zilin Chi,
Jiasheng Zhang,
Fan Cai,
Youzhi Zhu,
Dianping Tang,
Qingqiang Lin
Affiliations
Yan Ye
Fujian Normal University, College of Life Sciences, Fuzhou, Fujian, P. R. China
Yao Lin
Cooperation Base of Traditional Chinese Medicine-Oriented Chronic Disease Prevention and Treatment, Innovation and Transformation Center, Fujian University of Traditional Chinese Medicine, Fuzhou, China
Zilin Chi
Fujian Normal University, College of Life Sciences, Fuzhou, Fujian, P. R. China
Jiasheng Zhang
The First Affiliated Hospital of Fujian Medical University, Department of Thyroid and Breast Surgery, Fuzhou, Fujian, P. R. China
Fan Cai
Fujian Normal University, College of Life Sciences, Fuzhou, Fujian, P. R. China
Youzhi Zhu
The First Affiliated Hospital of Fujian Medical University, Department of Thyroid and Breast Surgery, Fuzhou, Fujian, P. R. China
Dianping Tang
Fuzhou University, Key Laboratory for Analytical Science of Food Safety and Biology (MOE & Fujian Province), State Key Laboratory of Photocatalysis on Energy and Environment, Department of Chemistry, Fuzhou, Fujian, P. R. China.
Qingqiang Lin
Fujian Normal University, College of Life Sciences, Fuzhou, Fujian, P. R. China
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.
