Method for detecting TP53 gene point mutation and kit for method

Abstract

The invention belongs to the field of gene mutation detection, and particularly relates to a TP53 gene point mutation detection method with high sensitivity and a kit for the method. The method comprises the following steps of: 1) extracting DNA according to the conventional technology; 2) then configuring a polymerase chain reaction (PCR) reaction system containing a specific primer pair, DNA polymerase and a PCR conventional component; and 3) performing PCR amplification on the DNA obtained in the step 1), analyzing the PCR product, and judging whether the PCR product has mutation, wherein the last second base of a sense primer in the specific primer pair is a sulfo modified base, and the DNA polymerase is high-fidelity DNA polymerase Pfu. The method has the advantages of rapidity, high specificity, high reliability, sensitivity, low condition dependency, wide application platform and the like.

Description

technical field [0001] The invention belongs to the field of gene mutation detection, and in particular relates to a highly sensitive TP53 gene point mutation detection method and a kit thereof. Background technique [0002] The TP53 gene (gene), discovered in 1979, is one of the genes most strongly associated with tumors so far. The gene is located on chromosome 17, and its protein product has the function of transcription factor. The genes controlled by the TP53 protein are involved in the process of cell division and survival. Like other tumor suppressor genes, the function of the TP53 protein is to prevent uncontrolled cell growth. [0003] The TP53 protein interacts directly with DNA, as well as with other proteins that govern how cells behave. When DNA damage or other cell damage is detected, TP53 immediately initiates the mechanism of apoptosis (apoptosis). The TP53 protein plays a key role in maintaining normal cell regulation, and about half of all tumors, regar...

AI Technical Summary

Problems solved by technology

Although the technology in method (1) is more sensitive than the existing mutation detection method, its required inactive primers are expensive, and the sensitivity is still low. It often takes 40-50 PCR cycles for a single copy of DNA molecules, and this method is not effective for There is a phenomenon of sequence discrimination in mutation recognition, that is, the pyrophosphate hydrolysis activity of Taq enzyme cannot hydrolyze some specific sequences, so it is difficult to bind inert primers, so there is no or little DNA polymerization reaction coupled with them

Although the method (2) has the characteristics of high sensitivity and reliability, the next-generation sequencing technology has yet to mature technically, and it relies on factors such as expensive equipment and a long detection process, which is still limited to experimental research and has not been practical Applied to clinical diagnosis

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