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Primer and method for simultaneously detecting XRCC1, ERCC1 and GSTP1 gene polymorphisms

A gene polymorphism, -TCTGACTCCCCTCCAGATT-3 technology, applied in biochemical equipment and methods, microbe measurement/inspection, DNA/RNA fragments, etc., can solve the problems of lack of specificity and high sensitivity, and achieve good accuracy, High sensitivity and improved detection efficiency

Inactive Publication Date: 2015-10-21
GUANGZHOU KINGMED DIAGNOSTICS CENT
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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 for detecting XRCC1 / ERCC1 / GSTP1 gene polymorphisms with good specificity, high sensitivity, and simultaneous detection of multiple sites

Method used

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  • Primer and method for simultaneously detecting XRCC1, ERCC1 and GSTP1 gene polymorphisms
  • Primer and method for simultaneously detecting XRCC1, ERCC1 and GSTP1 gene polymorphisms
  • Primer and method for simultaneously detecting XRCC1, ERCC1 and GSTP1 gene polymorphisms

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Primers

[0024] The inventor designed a large number of primers for the gene polymorphism sites of XRCC1 / ERCC1 / GSTP1, and through optimization and comparison of primer reaction conditions, primers with good specificity, no cross-reaction and very close PCR reaction conditions were screened out . The primers provided by the present invention include PCR amplification primers and SNaPshot PCR primers, and the PCR amplification primers and SNaPshot PCR primers are corresponding. All primer sequences provided by the present invention are compared through UCSC database, and there is no known SNP site.

[0025]

Embodiment 2

[0026] The specificity of embodiment two primers

[0027] The primers provided by the present invention were blasted in UCSC. The amplified fragment of XRCC1_R399Q primer was located at chr19: 44055651-44055888, with a length of 238bp; the amplified fragment of ERCC1_C118T primer was located at chr19: 45923504-45923722, with a length of 219bp; chr11: 67352602-67352777, the length is 176bp; the result is as follows Figure 1 to Figure 2 As shown, the amplified fragments of all primers covered the corresponding detection sites, and there were no other homologous genes.

[0028] 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 each primer were consistent with the reference sequence of the XRCC1 / ERCC1 / GSTP1 gene. The results are as follows Figure 3 to Figure 4 shown. Using the SNaPshot PCR primers in Table 1, the SNaPshot method is used for detection, ...

Embodiment 3

[0036] Example 3 Specificity of the method for detecting XRCC1 / ERCC1 / GSTP1 gene polymorphism

[0037] The specificity of this assay is defined as the negative coincidence rate. In this test, the detection of XRCC1_R399Q G / G genotype is defined as a negative result, the detection of ERCC1_C118T C / C genotype is defined as a negative result, and the detection of GSTP1_I105V A / A genotype is defined as a negative result.

[0038] The SNaPshot method provided by the present invention was used to detect 19 samples, and at the same time, the ARMS method or Sanger sequencing method was used for verification. The detection results of the SNaPshot method are consistent with those of the ARMS method or the Sanger sequencing method, as shown in Table 4. The specificity of this detection method is 100%.

[0039]

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Abstract

The invention belongs to the technical field of biological detection, and provides a primer for simultaneously detecting XRCC1, ERCC1 and GSTP1 gene polymorphisms. The primer comprises a PCR amplification primer and a SNaPshot PCR primer. The primer can achieve specific detection on XRCC1, ERCC1 and GSTP1 gene polymorphisms, causes no cross reaction and is good in accuracy.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a primer and a method for simultaneously detecting XRCC1, ERCC1 and GSTP1 gene polymorphisms. Background technique [0002] Platinum-based anticancer drugs, including cisplatin, carboplatin, and oxaliplatin, are currently clinically used anticancer drugs with high activity against a variety of cancers. They mainly eliminate cancer cells by causing intracellular DNA damage. [0003] X-ray repair cross-complementation group 1 (XRCC1), located on human chromosome 19q13.2-13.3, is 33 kb in size. Excision repair cross-complementation gene 1 (ERCC1), a human DNA damage repair gene, is located on chromosome 19q13.2-13.3. Glutathione S-transferase P1 (GSTP1), glutathione S-transferase (GST) is the most important phase II metabolic enzyme in the body, which exerts detoxification function by directly binding chemical carcinogens or by binding to glutathione , can...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6886C12Q1/6858C12Q2600/106C12Q2600/156C12Q2600/16C12Q2537/143C12Q2565/125
Inventor 赵薇薇胡昌明燕启江郭周萍
Owner GUANGZHOU KINGMED DIAGNOSTICS CENT
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