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Osteoarthritis diagnosing product and application thereof

A technology for osteoarthritis, products, applied in the field of biomedicine

Active Publication Date: 2016-09-21
范彧
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although various drug therapies have been used to treat the disease, they are ineffective for long-term control and prevention

Method used

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  • Osteoarthritis diagnosing product and application thereof
  • Osteoarthritis diagnosing product and application thereof
  • Osteoarthritis diagnosing product and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Example 1 High-throughput sequencing screening of differentially expressed genes

[0073] 1. Sampling

[0074] From October 2012 to December 2015, patients with osteoarthritis who were treated in the Department of Orthopedics of Peking Union Medical College Hospital were collected. A total of 10 cases were collected as the case group. A total of 4 cases were collected from patients with other diseases who were hospitalized in the Department of Orthopedics during the same period as controls. The synovial fluid samples of all subjects were obtained, numbered and stored in a -80°C low-temperature refrigerator.

[0075] The patients in the case group were all in line with the diagnostic criteria of knee OA and underwent knee arthroplasty; the control group were patients with meniscus injury and cruciate ligament injury who underwent arthroscopic surgery. According to the 1995 American Rheumatology Association diagnostic criteria for osteoarthritis, a patient with severe os...

Embodiment 2

[0097] Example 2 RT-PCR verification of CRNN gene expression in osteoarthritis patients and control synovial fluid

[0098] 1. Materials

[0099] The cartilage tissues of 42 cases of osteoarthritis patients and 8 cases of control cartilage tissues were selected and grouped and numbered. The patients in the case group were all in line with the diagnostic criteria of knee OA and underwent knee arthroplasty; the control group were patients with meniscus injury and cruciate ligament injury who underwent arthroscopic surgery.

[0100] 2. Method

[0101] 2.1 Carry out total RNA extraction to synovial fluid, with the extraction method of embodiment 1.

[0102] 2.2 Synthesis of cDNA by reverse transcription

[0103] use III Reverse Transcriptase (invitrogen, Cat. No. 18080-044) was used for cDNA reverse transcription, and the experimental operation was carried out according to the product manual. The specific operation was as follows:

[0104] Using a reverse transcription kit, ...

Embodiment 3

[0123] Example 3 Gene chip verification of CRNN gene expression in synovial fluid of patients with osteoarthritis and controls

[0124] 1, the acquisition of material, with embodiment 2.

[0125] 2, the extraction of total RNA, with the method for embodiment 1.

[0126] 3. Gene chip verification

[0127] After the total RNA was linearized and amplified, the cy3-UTP labeled and fluorescently labeled cRNAs were purified using the RNEASY Mini Kit, and the labeled cRNAs were fragmented with Amhion’s RNA Fragmentation Reagents. Human whole gene expression profile chips (4x 44K genes) from Agilent Company of the United States were used, hybridized in a chip hybridization oven at 65°C for 17 hours, then eluted, stained, and finally scanned with an Agilent DNA MicroarrayScanner scanner.

[0128] After the hybridized chip reads the data points by the chip scanner, the data is imported into the analysis software, and the genes whose absolute value of the natural logarithm of the ratio...

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Abstract

The invention discloses an osteoarthritis diagnosing product; the product can diagnose osteoarthritis through detecting the expression level of a CRNN gene in a biological sample, or a CRNN gene expression protein and fragments, analogs and derivatives thereof. The invention further discloses an application of the product in diagnosis of osteoarthritis. The invention discloses the gene CRNN related with osteoarthritis, and down-regulated expression of the CRNN gene or the gene expression protein and fragments, analogs and derivatives thereof in osteoarthritis tissues is further confirmed. With use of the gene for detection of osteoarthritis, early detection can be rapidly and effectively done, and a therapeutic target and an important basis are provided for gene therapy, drug therapy and other clinical applications.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a product for diagnosing osteoarthritis and its application. Background technique [0002] Osteoarthritis (OA) is a chronic disease in which the articular cartilage, which coats the ends of bones and forms the articular surfaces of joints, gradually deteriorates over time. A number of factors have been reported to predispose patients to osteoarthritis, including genetic predisposition, obesity, accident or sports trauma, surgery, medications, and strenuous demands. Osteoarthritis is thought to begin with damage to the articular cartilage. The two most common injuries to joints are sports-related injuries and long-term "repeated use" joint injuries. The joints most often affected by osteoarthritis are the knees, hips, and hands. In most cases, osteoarthritis in the knees and hips is more disabling than hand osteoarthritis due to the essential weight-bearing functions of these joints....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/68
CPCC12Q1/6883C12Q2600/158G01N33/6854G01N33/6893G01N2800/105
Inventor 范彧林进翁习生邱贵兴金今钱文伟陈俊马培杨跃梅
Owner 范彧
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