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Osteoarthritis disease screening genes PRXL2A and ACTR8 and use thereof

A technology for osteoarthritis and genes, applied in the field of disease diagnosis, can solve the problem of no molecular markers found

Active Publication Date: 2019-04-16
THE SECOND HOSPITAL OF HEBEI MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Osteoarthritis and rheumatoid arthritis are both systemic diseases, both large and small joints can be affected, but the two are two different diseases that must be distinguished, but no very good molecular markers that can distinguish the two have been found so far

Method used

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  • Osteoarthritis disease screening genes PRXL2A and ACTR8 and use thereof
  • Osteoarthritis disease screening genes PRXL2A and ACTR8 and use thereof
  • Osteoarthritis disease screening genes PRXL2A and ACTR8 and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 High-throughput sequencing of OA synovial tissue and normal synovial tissue

[0038] In our hospital, the synovial tissues of 3 patients with comminuted fractures were used as controls, and the synovial tissues of 4 patients with osteoarthritis were used as the case group. RNA was extracted from the samples to construct an mRNA library, and then the second-generation high Throughput sequencing technology to sequence mRNA. The data processing is completed through processes such as joint removal, low quality removal, and decontamination to obtain sequencing data.

[0039] Quantification of mRNA expression and differential expression analysis were performed using software cuffquan and cuffdiff. cuffquant is a tool specially used in the Cufflinks suite for expression evaluation and standardization. cuffdiff is a tool used in the Cufflinks suite to calculate differential expression. cuffdiff uses the quantitative results of cuffquant to compare the expression leve...

Embodiment 2

[0041] Example 2 Detection of PRXL2A gene and ACTR8 expression levels in OA and normal population

[0042] 1. Sample collection and extraction

[0043] For sample collection, 25 cases of peripheral blood from patients diagnosed with osteoarthritis in our hospital were selected, and 33 cases of peripheral blood from healthy people were selected as controls. RNA in serum was extracted using QIAGEN Blood RNA Extraction Kit. For specific steps, refer to the instruction manual.

[0044] 2. Reverse transcription:

[0045] 1) Mix 10pg-1μg total RNA template with 2μl 10× buffer, 2μl dATP (10mM), 0.5μl polyA polymerase, 0.5μl RNase inhibitor and RNase free water , the final volume was 20 μl, and incubated at 37°C for 1h.

[0046] 2) Add 1 μl of 0.5 μg / μl Oligo(dT) specific RT primer to the reaction tube, and incubate at 70° C. for 5 minutes.

[0047] 3) Immediately incubate on ice for at least 2 minutes to break the secondary structure of RNA and primers.

[0048] 4) Mix the above ...

Embodiment 3

[0069] Example 3 PRXL2A gene and ACTR8 differentiate RA and OA

[0070] The method for evaluating the efficacy of a single molecule or a diagnostic model is to establish a receiver operating characteristic (ROC) curve, and judge the ability of diagnosis by calculating the area under the curve (Area Under Curve). The value of the area under the ROC curve is between 1.0 and 0.5. In the case of AUC>0.5, the closer the AUC is to 1, the better the diagnostic effect. When the AUC is 0.5-0.7, the accuracy is lower, and when the AUC is 0.7-0.9 There is a certain accuracy when the AUC is above 0.9 and the accuracy is relatively high.

[0071] Clinically, arthritis can be divided into osteoarthritis, rheumatoid arthritis, ankylosing arthritis, reactive arthritis, gouty arthritis, etc. according to the etiology. Osteoarthritis and rheumatoid arthritis are both systemic diseases , both large and small joints can be affected, but the two are two different diseases, and there is no molecul...

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Abstract

The invention discloses osteoarthritis disease screening genes PRXL2A and ACTR8 and use thereof and particularly relates to the osteoarthritis disease screening genes PRXL2A and ACTR8 and the use thereof in preparation of osteoarthritis detection products. An inventor discovers molecular markers PRXL2A and ACTR8 closely related to osteoarthritis, and results show that the molecular markers have certain guide significance for the diagnosis of the osteoarthritis. In addition, the markers can also distinguish between osteoarthritis and rheumatoid arthritis to enable joint diagnosis of the two toachieve a superimposed effect. The genes PRXL2A and ACTR8 are expected to be candidate markers for molecular diagnosis of the osteoarthritis and for distinguishing between the osteoarthritis and the rheumatoid arthritis in clinical or health screening.

Description

technical field [0001] The present invention relates to the field of disease diagnosis. More specifically, the present invention relates to screening genes PRXL2A and ACTR8 for osteoarthritis (OA) disease and their application in the preparation of osteoarthritis detection products. The present invention also relates to the difference between the two in the preparation of Applications in Osteoarthritis and Rheumatoid Arthritis (RA) detection products. Background technique [0002] Osteoarthritis (OA) is a chronic joint disease characterized by degeneration of articular cartilage and secondary bone hyperplasia. The clinical diagnosis of osteoarthritis is mainly based on imaging examinations: X-ray findings are mainly joint space narrowing, subchondral bone sclerosis and cystic degeneration, osteophyte formation on the joint margin, articular surface collapse, deformation and joint subluxation etc.; MRI can show early cartilage, meniscus and other joint structure lesions, whi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12N15/11G01N33/68
CPCC12Q1/6883C12Q2600/136C12Q2600/158G01N33/6893G01N2800/102
Inventor 孙超田玉齐晅
Owner THE SECOND HOSPITAL OF HEBEI MEDICAL UNIV
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