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Method for detecting gene mutation

A detection method and gene mutation technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc.

Inactive Publication Date: 2012-07-04
辽宁琦润生物科技有限公司
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Problems solved by technology

[0009] The purpose of the present invention is to provide a method for detecting gene mutations, to solve the problem of complex equipment used in previous detection methods, high cost, and the possibility of false positives The problem

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  • Method for detecting gene mutation
  • Method for detecting gene mutation
  • Method for detecting gene mutation

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Embodiment 1

[0036] The P53 gene is named after encoding a protein with a molecular weight of 53 kDa, and it is an anti-cancer gene. Once the p53 gene is mutated, the P53 protein is inactivated, cell division is out of control, and cancer will occur. About half of human cancers are caused by mutations in this gene. Through the analysis of a large number of mutants in tumors, there are many P53 gene mutation sites in human tumors, there are hundreds of them, and the frequency of mutations in different tumors is different, among which the mutations at sites 175, 248, 249, 273, and 282 are the highest. Therefore, when detecting P53 gene mutations, it is very necessary to detect multiple mutation sites at the same time. We further explain the present invention by simultaneously detecting the gene mutations at the above five sites.

[0037] First, we use bioinformatics to know that the mutations at 175, 248, 249, 273, and 282 are p.R175H c.524G>A, p.R248W c.742C>T, p.R249S c.747G>T, p. .R273C...

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Abstract

The invention discloses a method for detecting gene mutation. Aiming at the defects of an allele-specific primer polymerase chain reaction (PCR) amplification method, recombinant plasmids containing wild type and mutant type genes to be detected are established respectively in advance by a molecular cloning technology, and two specific primers of which basic groups are specifically complementary with those of a 3'-tail-end mutation site or a wild site respectively are screened and determined by using the recombinant plasmids as a positive template, and the screened sequences of the specific primers are a key element of a detection system. A quantification standard curve is made by using the recombinant plasmids as the positive template, so that a specimen to be detected can be accurately quantified. A product can be used for a fluorescence ration PCR instrument of any model, and the cost of a reagent is equal to that of a fluorescence ration PCR reagent on the market currently, so that the defects of the allele-specific primer PCR amplification method can be overcome.

Description

technical field [0001] The invention relates to the field of gene detection, and in particular provides a method for detecting gene mutation. Background technique [0002] Mutations of human oncogenes or tumor suppressor genes are often early events in tumorigenesis. Detection of their mutations can detect tumors early, so it has very important clinical significance. At present, the detection of gene mutations generally includes sequencing method; gene chip hybridization method; PCR-RFLP method; PCR amplification product capillary electrophoresis analysis method; PCR high resolution melting curve analysis technology; PCR fluorescence quantitative probe method; allele specific Primer PCR amplification method, etc., these methods can detect gene mutations to varying degrees, but one of their common shortcomings is that they only qualitatively analyze mutants, and lack accurate quantification of mutants. In addition, these methods have more or less disadvantages such as high i...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 高劲松张英杰
Owner 辽宁琦润生物科技有限公司