CRSPR-cas13a driven RNA rapid detection method based on double-enzyme signal amplification strategy

Abstract

The invention discloses a CRSPR-cas13a driven RNA rapid detection method based on double-enzyme signal amplification strategy in the technical field of biological detection. The RNA rapid detection method based on the double-enzyme signal amplification strategy comprises the following steps: collecting a sample; treating a sample; assembling a magnetic separation probe; amplification and separation of primary enzymatic signal specifically triggered by a target sequence; amplification of secondary enzymatic signal and sending the signal; sending the detection signal of the target sequence. According to the present invention, the nucleic acid detection sequence is controllable, a suitable leader sequence can be designed according to the actual requirement so as to control the identified target sequence, which has extremely high specificity, extremely high sensitivity and good color development effect, and the detection results can be judged by naked eyes without relying on analytical instruments, the separation speed is high, the requirement of instrument is simple, and can be used to treat multiple groups of samples simultaneously, so that the manpower and the cost are saved.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a CRSPR-cas13a-driven RNA rapid detection method based on a dual-enzyme signal amplification strategy. Background technique [0002] RNA can be used as an effective biomarker for disease diagnosis. However, in the early stage of disease onset or incubation period, the RNA content in body fluids is low, which puts forward extremely high requirements for the sensitivity of the detection method. The classic rt-PCR detection method is time-consuming and laborious, relies on precise equipment such as fluorescent quantitative PCR instruments, and has a certain probability of false positives. In recent years, a highly sensitive detection method based on CRSPR-CAS13a has been proposed. However, the existing method still relies on fluorescent quantitative PCR instrument and other equipment. The operation process is complicated and the cost is high, which greatly affects the ...

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a CRSPR-cas13a-driven RNA rapid detection method based on a dual-enzyme signal amplification strategy to solve the problem of relatively complex biological fluid components and free nucleic acids that may interfere with the detection. Clinically available biological fluids are limited and the RNA content is low, and the concentration of purified nucleic acid sequences is difficult to reach the detection range

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