Construction of microRNA-21 terahertz metamaterial sensing method based on duplex-specific nuclease triggered rolling circle amplification
10.3760/cma.j.cn112150-20201111-01359
- VernacularTitle:基于双链特异性核酸酶触发滚环扩增的microRNA-21太赫兹超材料传感方法的构建
- Author:
Xinyu ZHAN
1
;
Sha YANG
;
Yang ZHANG
;
Xiang YANG
;
Weiling FU
Author Information
1. 陆军军医大学西南医院检验科，重庆 400038
- Keywords:
Duplex-specific nuclease;
Rolling circle amplification;
Terahertz metamaterial biosensor;
miRNA detection
- From:
Chinese Journal of Preventive Medicine
2021;55(2):212-218
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct of a terahertz metamaterial sensing method for detection of microRNA-21 (miRNA-21) based on duplex-specific nuclease triggered rolling circle amplification.Methods:First, a THz metamaterial sensing method was constructed, and verified by polyacrylamide gel electrophoresis and zeta potential; after optimizing the detection conditions of the sensor, different concentrations of miRNA-21 and other different miRNAs were detected. And this biosensor was compared with other miRNA detection methods; finally, the recovery rate of the biosensor was evaluated.Results:Under the optimal experimental conditions, through the dual signal amplification strategy of duplex-specific nuclease (DSN) cycle recognition and rolling circle amplification (RCA), the THz metamaterial sensor has a response range of 10 fmol/L to 10 nmol/L to the target miRNA-21, with a detection limit of 8.49 fmol/L. And the biosensor has good specificity with the ability to recognize the target miRNA-21 from a variety of microRNAs. And the experiment with recovery rate from 94.33% to 115.33% has been further verified in commercial human serum samples.Conclusion:The terahertz biosensor can achieve highly sensitive and specific detection of the target miRNA-21, which proves the potential for label-free diagnosis and early warning of miRNA-related diseases.
