Multiple nucleic acid detection system as well as preparation method and application thereof

Abstract

The invention relates to a multiple nucleic acid detection system, which comprises an amplification primer group and a detection probe group aiming at a target nucleic acid sequence, the detection system comprises a Target probe modified by LNA and a Beacon probe for achieving a fluorescence quenching effect through 1-8 continuous G basic groups, and meanwhile, based on the proposal of the multiple nucleic acid detection system, the inventor combines a touchdown PCR (Polymerase Chain Reaction) program to detect the target nucleic acid sequence. The invention further provides a multiple nucleic acid detection method which can further reduce non-specific amplification in PCR reaction and improve detection sensitivity. Therefore, the limitation of traditional real-time fluorescent quantitative PCR typing is overcome, single-tube multi-typing is realized through special signal and melting curve analysis, and accurate qualitative detection of each target gene in at most 20 to-be-detected target nucleic acid sequences in a sample can be realized with a simpler reaction system and lower detection cost.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a multiple nucleic acid detection system and its preparation method and application. Background technique [0002] Real-time fluorescent quantitative PCR method is a commonly used nucleic acid detection method in molecular biology. Compared with ordinary PCR method, it is easy to operate and widely used. During the PCR amplification process of the target gene of the sample to be tested, the fluorescent signal generated by the reaction system is monitored in real time by the instrument, and the PCR process is detected in real time. Ordinary PCR amplification technology requires electrophoresis analysis of the product after the amplification is completed. The analysis process is cumbersome and time-consuming. At the same time, the opening of the PCR product may cause contamination of the laboratory environment. [0003] The methods for realizing fluorescent signals are mainly divided in...

AI Technical Summary

Problems solved by technology

However, when the method described in this patent application is used to implement multiplex real-time PCR that needs to distinguish each target sequence, three probes need to be designed for each target sequence, which leads to more complex reaction system and high detection cost

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