Multi-quenching fluorescent probe and method for detecting target nucleic acid sequence

Abstract

The invention discloses a multi-quenching fluorescent probe and a method for detecting a target nucleic acid sequence. The multi-quenching fluorescent probe has two or more quenching groups. The method for detecting the target nucleic acid sequence comprises steps as follows: a to-be-detected nucleic acid sequence is taken as a template, a fluorescent PCR reaction is performed under the action ofa corresponding primer and the multi-quenching fluorescent probe, and a dissolution curve is drawn. Compared with other dissolution curve analysis technologies, the target nucleic acid sequence detection method based on cutting of hybrid sequence fluorescence quenching groups has the advantages that when no target sequence is detected, the fluorescent probe does not produce a fluorescent signal, so that the background is notably reduced during analysis of the dissolution curve, and detection of other positive target sequences cannot be affected; when multiple fluorescent probes are detected, the background signals of the fluorescent probes cannot interfere with each other, and accordingly, interpretation of multiple PCR is clearer.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering. More specifically, the invention relates to a multiple quenching fluorescent probe and a detection method for target nucleic acid sequence detection based on hybridization sequence fluorescence quenching group cleavage and melting curve analysis. Background technique [0002] Fluorescent PCR technology is a common technique for detecting target nucleic acid sequences. Currently, commonly used fluorescent PCR detection methods include Taqman probe method, molecular beacon probe method, hybridization probe method, etc. Among them, the Taqman probe method mainly relies on the exonuclease activity of nucleic acid polymerase. During the PCR extension process, the fluorescent probe hybridized with the target sequence is cut, and the 5' end fluorescent group is released. After the 5' end fluorescent group is released, The resulting rise in fluorescence detects the presence of the target sequ...

AI Technical Summary

Problems solved by technology

[0004] At present, when the fluorescent probe melting curve technology uses multiple fluorescent probes in the same fluorescent channel for detection, even if the fluorescent probes are not combined with the target sequence, the probes themselves or between the probes will entangle and mishybridize under low temperature conditions. Background signal, the background signals of multiple fluorescent probes interfere with each other and superimpose, resulting in an excessively high fluorescent background, which affects the interpretation of the melting curve

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