Gene mutation detection kit based on high-resolution melting curve of non-fluorescence labeled probe, detection method and application of gene mutation detection kit

Abstract

The invention relates to the technical field of biomedicine, in particular to a gene mutation detection kit based on a high-resolution melting curve of a non-fluorescence labeled probe, a detection method and application of the gene mutation detection kit. The kit comprises a system composed of a non-fluorescence labeled probe, two forward and reverse amplification primers, a DNA polymerase with 5 '-3' exonuclease activity deletion, a fluorescent dye, a dNTP mixed solution and a PCR buffer solution. Wherein the non-fluorescence labeled probe is a non-fluorescence labeled probe of which the 3'end is blocked by group modification and thus loses the ability to extend. According to the detection system, common oligonucleotide is used for replacing an expensive fluorescent probe for nucleic acid recognition, the purpose of gene polymorphism detection is achieved, a non-fluorescence labeled probe is combined with a target sequence with a mutation site, and sequences at different melting temperatures are converted into signal generation through melting curve analysis, so that rapid detection of gene mutation is realized through the non-fluorescence labeled probe.

Description

technical field [0001] The invention relates to the field of biomedical technology, in particular to a gene mutation detection kit, detection method and application thereof based on a high-resolution melting curve of a non-fluorescent labeled probe. Background technique [0002] The study of gene mutation has become one of the hotspots in life science research, and the detection methods have also developed rapidly. The mutation types of human cell oncogenes have been mentioned above. For the detection of gene mutations, before 1985, the Southern blot method could be used to screen out gene mutations such as deletion, insertion, and frameshift recombination. For mutations that cannot be detected by this method, complex and time-consuming DNA sequence analysis methods can only be applied. Polymerase chain reaction (polymerase chain reaction PCR) technology is the most significant progress in mutation research, which has made great progress in gene mutation detection technolog...

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Problems solved by technology

[0006] The present invention aims to overcome the defects of high instrument price, difficult operation, certain false negatives and false positives, high detection cost, low clinical popularity and inability to detect clinical specimens on a large scale in the existing methods for detecting gene mutations, and provides A gene mutation detection kit, detection method and its application based on the high-resolution melting curve of non-fluorescent labeled probes are developed to solve the problems of complicated operation, high cost and easy false positive in previous detection methods

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