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Application of XRCC1 gene polymorphism to rheumatoid arthritis diagnostic effectiveness

A technology for arthritis and rheumatoids, applied in the field of medicine and biology, can solve the problems of no related reports and achieve good sensitivity

Active Publication Date: 2017-11-28
THE SECOND AFFILIATED HOSPITAL TO NANCHANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

XRCC1 is an important component in the base repair system, and the single nucleotide polymorphism in its conserved region will affect the function of XRCC1 protein and improve the DNA repair function, but the current research on the relationship between XRCC1 single nucleotide polymorphism and rheumatoid arthritis Inflammatory relationship No relevant reports

Method used

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  • Application of XRCC1 gene polymorphism to rheumatoid arthritis diagnostic effectiveness
  • Application of XRCC1 gene polymorphism to rheumatoid arthritis diagnostic effectiveness
  • Application of XRCC1 gene polymorphism to rheumatoid arthritis diagnostic effectiveness

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1 Determination of the Relationship Between Gene Variability and Rheumatoid Arthritis

[0021] 1. Research object

[0022] Whole blood samples from 300 patients with clinically confirmed rheumatoid arthritis after hospitalization in XXX Hospital since August 2015 (sodium citrate anticoagulant 3ml). All whole blood samples were separated from plasma within 2 hours after collection, centrifuged at 4000rmp for 10 minutes, 1ml of plasma was collected in 1.5ml EP tube, and the remaining whole blood was divided into 2 tubes. Aliquoted samples were stored at -40°C. This study was approved by the Hospital Ethics Committee, and all subjects or their family members gave informed consent.

[0023] Genomic DNA was extracted using a blood extraction kit. The sequences of primers for PCR amplification are SEQ ID NO: 1 and 2. The reaction system is 20 μL: the concentration of each primer is 0.3 μM, 2.5 mM Mg+, 0.2 mM dNTP, 1×PCRBuffer, 1 μL Taq enzyme, 0.5 μL template, and...

Embodiment 2

[0027] Example 2XRCC1-641 Genotyping Analysis

[0028] Get GG, GC and CC genotypes, use the PCR method of Example 1 to amplify respectively, and use restriction endonuclease to cut the amplified products respectively, the enzyme is HaeIII, and the enzyme cutting system is 10 microliters: PCR 3.5 microliters of the product, 1 microliter of 10× enzyme digestion buffer, 2 U of restriction endonuclease, filled to 10 microliters with sterilized three-distilled water. The reaction condition is 65°C water bath for 2-3h. After digestion, 2 microliters of the digested products were separated by 8% polyacrylamide gel electrophoresis. The results are shown in Table 2:

[0029] Table 2 Restriction endonuclease names and restriction enzyme fragments

[0030] PCR product

[0031]This is consistent with the genotype results obtained by sequencing, which fully demonstrates that the enzyme-cut typing of the present invention can be effectively used for genotype identification and ...

Embodiment 3

[0032] Example 3 Clinical Patient Identification

[0033] Take 10 arthritis patients and normal patients, and use the kit to extract the corresponding DNA. The sequences of primers for PCR amplification are SEQ ID NO: 1 and 2. The reaction system is 20 μL: the concentration of each primer is 0.3 μM, 2.5 mM Mg2+, 0.2 mM dNTP, 1×PCR Buffer, 1 μL Taq enzyme, 0.5 μL template, and the balance is ultrapure water. The amplification conditions were pre-denaturation at 96°C for 3min, denaturation at 95°C for 6sec, annealing at 60°C for 6sec, and extension at 72°C for 15sec for 35 cycles. EvaGreen Mix amplification conditions were pre-denaturation at 96°C for 3min, denaturation at 96°C for 45sec, annealing at 58.5°C for 45sec, and extension at 72°C for 50sec for 35 cycles. The amplified product was purified by a PCR purification kit.

[0034] The amplified products were respectively digested with restriction endonuclease, the enzyme was HaeIII, and the digestion system was 10 microli...

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Abstract

The invention belongs to the technical field of biological medicine and particularly relates to an application of XRCC1 gene polymorphism to rheumatoid arthritis diagnostic effectiveness and an application of SNP loci of the XRCC1 gene to preparation of a detection kit for diagnosing rheumatoid arthritis susceptibility. The correlation between polymorphism of the 641th lotus of the XRCC1 gene and rheumatoid arthritis is discovered for the first time, so that a method and a kit for detecting rheumatoid arthritis susceptibility risk through SNP are proposed on the basis, clinical experiment research verifies that the detection method has feasibility, and the detection kit has good sensitivity, stability and specificity.

Description

technical field [0001] The invention relates to the use of XRCC1 gene polymorphism in rheumatoid arthritis diagnosis and / or prognosis kit and the use of XRCC1 gene blocker in preparing medicine for treating rheumatoid arthritis, belonging to the field of medical biotechnology. Background technique [0002] Rheumatoid arthritis is a common acute or chronic inflammation of connective tissue. Rheumatoid arthritis should broadly include rheumatoid arthritis (RA), which can recur and involve the heart. Clinically, it is characterized by joint and muscle migratory soreness, heaviness, and pain. It belongs to an allergic disease and is one of the main manifestations of rheumatic fever. It usually starts with acute fever and joint pain. Rheumatoid arthritis is the manifestation of rheumatic fever in the joints. Its typical symptoms are migratory and polyarthritis. It is common to transfer from one joint to another. The local lesion is red, swollen, burning, and severe pain. Some pa...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6883C12Q2600/118C12Q2600/156C12Q2600/172
Inventor 邹叶青黄波付晓伟王小中
Owner THE SECOND AFFILIATED HOSPITAL TO NANCHANG UNIV
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