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

目的:构建一种太赫兹(THz)超材料传感方法用于microRNA-21(miRNA-21)的信号放大检测。方法:首先构建THz超材料传感方法,并用聚丙烯酰胺凝胶电泳及zeta电位验证方法的可行性;对传感器检测条件进行优化之后,对不同浓度的miRNA-21以及其他不同的miRNAs进行检测,并与其他microRNA检测方法进行比较;最后,对该传感器的回收率进行了评价。结果:在最优实验条件下,通过双链特异性核酸酶(DSN)循环识别与滚环扩增(RCA)的双重信号放大策略,该THz超材料传感器对靶标miRNA-21的响应范围为10 fmol/L至10 nmol/L,检测限为8.49 fmol/L。并且该传感器具有较好的特异性,具备了从多种miRNAs中识别靶标miRNA-21的能力,并且在商业化人血清样本中的回收率可达94.33%到115.33%。结论:该THz传感器可以实现靶标miRNA-21的高灵敏、高特异性检测,具备了在miRNA相关疾病无标记诊断与早期预警的潜力。

Construction of microRNA-21 terahertz metamaterial sensing method based on duplex-specific nuclease triggered rolling circle amplification

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.