基于CRISPR/Cas12a系统的比色检测方法探究

Study on Colorimetric Detection Based on Clustered Regularly Interspaced Short Palindromic Repeats/Cas12a System

基于Clustered Regularly Interspaced Short Palindromic Repeats/Cas12a(CRISPR/Cas12a)系统的特征基因识别检测技术,已经可以实现DNA的普适性检测。同时,基于CRISPR/Cas12a系统的反式酶切活性,可以降解(有目标DNA)或者不降解(无目标DNA)检测体系中添加的指示探针,设计一对通用的DNA纳米金探针进行裸眼可视化DNA比色检测。整个检测过程与结果读出不依赖于实时定量荧光PCR仪、恒温培养箱等大型仪器,整个流程可以在1小时之内完成。

The identification and detection technology of pathogenic microorganism characteristic genes based on the Clustered Regularly Interspaced Short Palindromic Repeats/Cas12a(CRISPR/Cas12a)system has been able to realize the universal detection of DNA pathogenic genes.At the same time,based on the trans digestion activity of CRISPR/Cas12a system,the indicator probes added in the detection system can be degraded(with target pathogenic microorganisms)or not degraded(without target pathogenic microorganisms),a pair of universal DNA nanogold probes are designed for naked-eye visual detection.The whole detection process and result reading do not depend on large-scale instruments such as real-time quantitative fluorescent PCR instrument and thermostatic incubator,and the whole process can be completed within 1 hour.