Primer and method for detecting XRCC1 gene polymorphism

A technology for gene polymorphism and amplification primers, which is applied in the field of primers for detecting XRCC1 gene polymorphisms, can solve the problems of lack of specificity and high sensitivity, and achieve the effects of good specificity, high sensitivity and improved detection efficiency

Inactive Publication Date: 2015-10-21
GUANGZHOU KINGMED DIAGNOSTICS CENT +1
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

The prior art lacks a primer and method thereof with good specif

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  • Primer and method for detecting XRCC1 gene polymorphism
  • Primer and method for detecting XRCC1 gene polymorphism
  • Primer and method for detecting XRCC1 gene polymorphism

Examples

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Comparison scheme
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Embodiment 1

[0019] Example 1 Primers

[0020] The inventor designed a large number of primers for the gene polymorphism site of XRCC1, and selected primers with good specificity through optimization and comparison of primer reaction conditions. 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.

[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 XRCC1_R399Q primer was located at chr19: 44055651-44055888, with a length of 238bp. The result is as 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 XRCC1 gene. The results were as follows: figure 2 shown. Using the SNaPshot PCR 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 with expec...

Embodiment 3

[0032] Example 3 Specificity of the method for detecting XRCC1 gene polymorphism

[0033] The specificity of this assay is defined as the negative coincidence rate. This test defines the detection of the XRCC1_R399Q G / G genotype as a negative result. The SNaPshot method provided by the present invention was used to detect 9 samples, and at the same time, the ARMS method was used for verification. The detection results of the SNaPshot method are consistent with those of the ARMS method, as shown in Table 4. The specificity of this detection method is 100%.

[0034]

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Abstract

The invention provides a primer for detecting XRCC1 gene polymorphism. The primer for detecting XRCC1 gene polymorphism comprises a PCR amplification primer and a SNaPshot PCR primer. The primer for detecting XRCC1 gene polymorphism belongs to the technical field of biological detection. The primer for detecting XRCC1 gene polymorphism can achieve the specific detection on the XRCC1 gene polymorphism, and the accuracy is good.

Description

technical field [0001] The invention belongs to the technical field of biological detection, in particular to a primer and a method for detecting XRCC1 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] X-ray repair cross-complementary group 1 (XRCC1), located on human chromosome 19q13.2-13.3, with a size of 33 kb, XRCC1 is the first mammalian gene isolated to affect the sensitivity of cells to ionizing radiation, and is widely involved in DNA damage repair. [0004] Existing research data show that the frequencies of XRCC1_R399Q R / R, R / Q and Q / Q genotypes are about 55%, 37%, and 8%, respectively. Therefore, the detection of XRCC1 gene polymorphism is helpful to predict the prognosis of platinum-based chemotherapy, and is ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6886C12Q1/6858C12Q2600/106C12Q2600/156C12Q2531/113C12Q2565/125
Inventor 赵薇薇胡昌明燕启江郭周萍
Owner GUANGZHOU KINGMED DIAGNOSTICS CENT
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