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Application of hsa-miR-17-5p as diagnostic marker of rheumatoid arthritis

A rheumatoid and arthritis technology, applied in the field of biomedicine, can solve problems such as low sensitivity and poor specificity of diagnostic markers, and achieve high sensitivity, good clinical application value, and good specificity

Active Publication Date: 2019-01-11
THE THIRD AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved in the present invention is to overcome the problems of poor specificity and low sensitivity of existing rheumatoid arthritis diagnostic markers, and to provide the application of serum hsa-miR-17-5p as a rheumatoid arthritis diagnostic marker, Provide a new target for the diagnosis of rheumatoid arthritis, and have good clinical application value for the diagnosis of rheumatoid arthritis

Method used

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  • Application of hsa-miR-17-5p as diagnostic marker of rheumatoid arthritis
  • Application of hsa-miR-17-5p as diagnostic marker of rheumatoid arthritis
  • Application of hsa-miR-17-5p as diagnostic marker of rheumatoid arthritis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 Detection of hsa-miR-17-5p level in synovial tissue of patients with rheumatoid arthritis by qRT-PCR

[0035] 1. Preparation of cDNA

[0036] The synovial tissue of patients with rheumatoid arthritis and the synovial tissue of bone and joint replacement patients were obtained from the Third Affiliated Hospital of Guangzhou Medical University. The synovial tissue was collected aseptically. After the operation, the synovial tissue of the joint was collected in a clean and sterile 50mL centrifuge tube, placed on ice, and the adipose tissue, bones, blood vessels, etc. were removed on the aseptic operating table, and then washed with PBS for 3 all over. RNA was extracted using miRNeasy Micro Kit from QIAGEN, and reverse transcribed into cDNA.

[0037] 2. PCR detection

[0038] Using cDNA as a template, according to the operating instructions of the miScript SYBR Green PCR kit (QIAGEN Company), with the primers shown in Table 1, qRT-PCR experiments were carried o...

Embodiment 2

[0044] Example 2 Detection of hsa-miR-17-5p level in serum of patients with rheumatoid arthritis by qRT-PCR

[0045] 1. Preparation of cDNA

[0046] Sera from patients with rheumatoid arthritis and healthy subjects were obtained from the Third Affiliated Hospital of Guangzhou Medical University. Serum was placed in a high-speed refrigerated centrifuge at 10,000 rpm for 10 minutes to remove cell debris. RNA was extracted using QIAGEN's miRNeasy Serum / PlasmaKit, and reverse-transcribed into cDNA.

[0047] 2. PCR detection

[0048] Using cDNA as a template, according to the operating instructions of the miScript SYBR GreenPCR kit (QIAGEN company), with the primers shown in Table 2, qRT-PCR experiments were carried out;

[0049]

[0050] PCR reaction parameters: 96°C for 3min, 60°C for 40s, 45 cycles. The above experiments were repeated three times, using cel-miR-39 as the internal control, and after 2 -△△CT Calculate the difference in the expression level of hsa-miR-17-5p...

Embodiment 3

[0052] Example 3 ROC curve analysis of the diagnostic function of hsa-miR-17-5p expression level in patients with rheumatoid arthritis

[0053] 1. Using SPSS17.0 software to analyze the diagnostic role of hsa-miR-17-5p in serum in patients with rheumatoid arthritis.

[0054] 2. Conclusion: The ROC curve is attached image 3 As shown in Figure A in the middle, the results show that the AUC area is 0.864, indicating that hsa-miR-17-5p in serum has a good value for the diagnosis of rheumatoid arthritis; ROC curve analysis, the results are attached image 3 As shown in Figure B in the middle, it shows that the sensitivity and specificity of hsa-miR-17-5p for the diagnosis of rheumatoid arthritis are 80% and 80% respectively; the 95% confidence interval is 0.751-0.977, which has good specificity and sensitivity high feature.

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Abstract

The invention discloses an application of hsa-MiR-17-5p as a diagnostic marker of rheumatoid arthritis. The application of hsa-MiR-17-5p as a diagnostic marker of rheumatoid arthritis firstly demonstrates that hsa-MiR-17-5p is overexpressed in synovial tissue of rheumatoid arthritis patients and secreted into serum, and hsa-MiR-17-5p level is increased significantly. Therefore, the present invention provides a novel diagnostic marker of rheumatoid arthritis, namely serum hsa-MiR-17-5p, which has good specificity and high sensitivity for rheumatoid arthritis, and overcomes the shortcomings of poor specificity and lower sensitivity of rheumatoid factor RF and anti-cyclic citrulline antibody ACPA, which are the existing diagnostic markers of rheumatoid arthritis, and has good diagnostic valuefor rheumatoid arthritis.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and more specifically relates to the application of hsa-miR-17-5p as a diagnostic marker for rheumatoid arthritis. Background technique [0002] Rheumatoid arthritis (RA) is an autoimmune disease, and its pathological features are inflammatory response and increased secretion of inflammatory cytokines. The clinical manifestations of RA are long-term chronic inflammatory infiltration, joint pain, joint swelling and joint deformity, which bring great harm to human health and life. Epidemiological studies have found that the incidence of rheumatoid arthritis in women is about 1.16% in the world, and 0.44% in men. At present, rheumatoid arthritis has become a public health problem to be solved urgently, and it is also a hot spot of clinical research. Studies have shown that early diagnosis and treatment of rheumatoid arthritis can effectively improve the quality of life and prognosis of patient...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/158C12Q2600/178
Inventor 洪宏海
Owner THE THIRD AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIVERSITY
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