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Primer for detecting ERCC2 gene polymorphism, and method thereof

A technology for gene polymorphism and amplification primers, which is applied in the field of primers for detecting ERCC2 gene polymorphisms, can solve the problems of lack of specificity and high sensitivity, achieve good specificity, high sensitivity, and improve detection efficiency

Inactive Publication Date: 2015-11-11
GUANGZHOU KINGMED DIAGNOSTICS CENT +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The prior art lacks a primer and method thereof with good specificity, high sensitivity, and the ability to detect polymorphisms of the ERCC2 gene

Method used

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  • Primer for detecting ERCC2 gene polymorphism, and method thereof
  • Primer for detecting ERCC2 gene polymorphism, and method thereof
  • Primer for detecting ERCC2 gene polymorphism, and method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1 Primer

[0020] The inventor designed a large number of primers for the gene polymorphism site of ERCC2, and screened out primers with good specificity through optimization and comparison of primer reaction conditions. The primers provided by the present invention include PCR amplification primers and SNaPshotPCR primers, and the PCR amplification primers and SNaPshotPCR primers are corresponding. All primer sequences provided by the present invention are compared through UCSC database, and there is no known SNP site.

[0021]

Embodiment 2

[0022] The specificity of embodiment two primers

[0023] The primers provided by the present invention were blasted in UCSC, and the amplified fragment of the ERCC2_L751Q primer was located at chr19:45854837+45855007, with a length of 171bp; the results were as follows figure 1 As shown, the amplified fragments of the primers all covered the corresponding detection sites, and there were no other homologous genes.

[0024] The PCR amplification primers in Table 1 were used to amplify and Sanger sequence the test samples respectively. The sequencing results showed that the fragments amplified by the primers matched the reference sequence of the ERCC2 gene. The results were as follows: figure 2 shown. Using the SNaPshotPCR primers in Table 1, the SNaPshot method is used for detection, and the results are as follows image 3 shown. from image 3 It can be seen that the relative position of each product peak and the bases incorporated in the sequencing reaction are in line wi...

Embodiment 3

[0032] Example 3 Specificity of the method for detecting ERCC2 gene polymorphism

[0033] The specificity of this assay is defined as the negative coincidence rate. This test defines the detection of ERCC2_L751QA / A genotype as a negative result. The SNaPshot method provided by the present invention was used to detect 19 samples, and at the same time, the Sanger sequencing method was used for verification. The detection results of the SNaPshot method are consistent with the results of the Sanger sequencing method, as shown in Table 4. The specificity of this detection method is 100%.

[0034]

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PUM

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Abstract

The invention provides a primer for detecting the ERCC2 gene polymorphism. The primer comprises a PCR amplification primer and an SNaPshot PCR primer. The invention belongs to the technical field of biological detection. The primer provided by the invention can realize specific detection of the ERCC2 gene polymorphism, and has good accuracy.

Description

technical field [0001] The invention belongs to the technical field of biological detection, in particular to a primer and a method for detecting ERCC2 gene polymorphism. Background technique [0002] Platinum-based anticancer drugs, including cisplatin, carboplatin, and oxaliplatin, are currently clinically used anticancer drugs with high activity against various cancers. They mainly eliminate cancer cells by causing intracellular DNA damage. [0003] Excision repair cross-complementation gene 1 (ERCC1), a human DNA damage repair gene, is located on chromosome 19q13.2-13.3. Excision repair cross-complementation gene 2 (ERCC2), involved in nucleotide excision repair and gene transcription, plays an important role in DNA damage repair. [0004] Existing research data show that ERCC1_C118TC / C, C / T, T / T genotype frequencies are about 52.6%, 43.1%, 4.2% respectively; ERCC2_L751QK / K, K / Q, Q / Q genotype frequencies are about 84.92% %, 15.08%, 0%. Therefore, the detection of ERCC...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6886C12Q2600/106C12Q2600/156
Inventor 赵薇薇胡昌明燕启江郭周萍
Owner GUANGZHOU KINGMED DIAGNOSTICS CENT
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