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A set of primers capable of simultaneously detecting ERCC4 and XRCC1 genetic polymorphism, and applications

A gene polymorphism, reverse primer technology, applied in recombinant DNA technology, microbial determination/inspection, biochemical equipment and methods, etc., can solve the problems of good accuracy, high specificity and sensitivity, and achieve high sensitivity, The effect of reducing amplification procedures and ensuring reliability

Inactive Publication Date: 2018-12-07
KUNMING UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Nowadays, with the assistance of a third-party testing company, single-gene sequencing methods are used to detect the polymorphisms of these sites respectively; there is a lack of a specificity, high sensitivity, good accuracy, and simultaneous realization of multiple sites. Detection techniques and methods

Method used

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  • A set of primers capable of simultaneously detecting ERCC4 and XRCC1 genetic polymorphism, and applications
  • A set of primers capable of simultaneously detecting ERCC4 and XRCC1 genetic polymorphism, and applications
  • A set of primers capable of simultaneously detecting ERCC4 and XRCC1 genetic polymorphism, and applications

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1: ERCC4rs1799801, XRCC1rs25487 Gene polymorphism SNaPshot detection

[0021] This detection method utilizes a set of disclosed simultaneous detection ERCC4rs1799801, XRCC1rs25487 The primers for gene polymorphism are amplified by multiplex PCR method to amplify the gene fragment where the SNP site to be detected is located, and the fluorescent label single base extension technology (SNaPshot) combined with capillary electrophoresis technology is used for gene polymorphism detection, and the detection results are applied to GENEMAPPER5 software Perform analysis, determine the SNP site corresponding to the extension product according to the shift position of the peak, and determine the genotype of the SNP site according to the fluorescence signal of the peak.

[0022] This detection method can be used to detect DNA samples derived from EDTA anticoagulated peripheral blood of lung cancer patients. After the multiplex PCR method is used to amplify the gene fragm...

Embodiment 2

[0045] Embodiment 2: detection of specificity, sensitivity, accuracy, precision

[0046] 1. Specific detection

[0047] The specificity of this detection method is defined as the negative coincidence rate. The present invention detects 20 examples of non-small cell lung cancer samples with the SNaPshot method, and the detection results show: ERCC4 rs17990801 The negative rate was 40%, XRCC1 rs25487 The negative rate was 60%, of which only 3 cases were negative for all sites, which was consistent with the test results of sanger sequencing (as shown in Table 4), and the specificity of the present invention was 100%.

[0048] Table 4 Detection method specificity

[0049] ;

[0050] 2. The sensitivity of the detection method

[0051] The sensitivity of this detection method is defined as the positive coincidence rate. The present invention detects 20 samples, and the SNaPshot method detects 17 samples with positive mutations, which are consistent with the detection results...

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Abstract

The invention discloses a set of primers capable of simultaneously detecting ERCC4 and XRCC1 genetic polymorphism. The primers include a PCR amplification primer and an SNaPshot PCR primer, and simultaneous detection loci include ERCC4 rs1799801 and XRCC1 rs25487; compared with conventional individual amplification performed on multiple loci, amplification programs can be minimized, amplificationcosts can be reduced, and high sensitivity, accuracy and precision can be guaranteed by applying the primers to the selection of clinical chemotherapy drugs, and therefore, the reliability of a methodcan be ensured; and the primers are helpful for doctors to correctly select drugs and treatment schemes for patients, so that the survival rates of cancer patients can be increased.

Description

technical field [0001] The invention belongs to the field of gene polymorphism detection, in particular to a group of simultaneous detection ERCC4 and XRCC1 Gene polymorphism primers and applications. Background technique [0002] The lethality rate of cancer is second only to cardiovascular and cerebrovascular diseases, and the clinical treatment method is based on platinum-based chemotherapy; the target of platinum-based drugs is DNA, which damages DNA through inter-strand interactions and slows down the malignant proliferation of tumor cells rate, while the single nucleotide polymorphism of DNA repair ability affects the prognosis of patients; nucleotide excision repair (NER), which is a multifunctional and complex DNA damage excision pathway, is initiated by recognizing DNA damage. The main steps in this process are unwinding of the double helix at the site of DNA damage, excision of the single-stranded DNA fragment containing the damage, repair of the gap and filling...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q1/6886C12Q2600/106C12Q2600/16C12Q2600/156C12Q2531/113C12Q2537/143C12Q2565/125
Inventor 刘宁唐文如罗瑛旦菊花盛苗苗张继虹贾舒婷吴晓明刘静谢晓丽周若宇
Owner KUNMING UNIV OF SCI & TECH
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