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

PeerJ

article

Yan Ye¹  Yao Lin²  Zilin Chi¹  Jiasheng Zhang³  Fan Cai¹  Youzhi Zhu³  Dianping Tang⁴  Qingqiang Lin¹ 

[1] Fujian Normal University, College of Life Sciences;Cooperation Base of Traditional Chinese Medicine-Oriented Chronic Disease Prevention and Treatment, Innovation and Transformation Center, Fujian University of Traditional Chinese Medicine;The First Affiliated Hospital of Fujian Medical University, Department of Thyroid and Breast Surgery;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

关键词: RCA;    Fluorescent detection;    Biosensor;    miR-129-2-3p;   

DOI  :  10.7717/peerj.14257

学科分类：社会科学、人文和艺术（综合）

来源: Inra

【 摘 要 】

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.

【 授权许可】

CC BY