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Application of circRNA in the preparation of products for diagnosing osteoarthritis

A technology of osteoarthritis and a kit, which is applied in the field of biomedicine, can solve the problems of ineffectiveness, and achieve the effect of rapid and effective early detection

Active Publication Date: 2021-03-05
PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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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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  • Application of circRNA in the preparation of products for diagnosing osteoarthritis
  • Application of circRNA in the preparation of products for diagnosing osteoarthritis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 High-throughput sequencing screening of differentially expressed genes

[0028] 1. Sampling

[0029] 19 cases of cartilage tissue samples (average age 64.5 years old, age range 58-75 years old, 2 males, 17 females) were obtained from the osteoarthritis operation in the Department of Orthopedics of Peking Union Medical College Hospital. All samples were confirmed by pathological examination and diagnosed as knee joint Osteoarthritis; normal cartilage tissue comes from the Department of Orthopedics, Peking Union Medical College Hospital, which is the articular cartilage tissue of patients undergoing trauma surgery. The obtained tissues were stored in a -80°C low-temperature refrigerator after serial numbering. All clinical samples used in this study were informed to the patients and approved by the Ethics Committee of our hospital.

[0030] 2. Total RNA extraction from tissue samples

[0031] use Reagent (Invitrogen, Carlsbad, CA, USA) was used to extract t...

Embodiment 2

[0056] Example 2 RT-PCR verification of the expression of hsa_circ_0045714 and hsa_circ_0005567 in cartilage tissue of OA patients

[0057] 1. Materials

[0058] Seventeen osteoarthritis cartilage tissue samples were collected from Peking Union Medical College Hospital for osteoarthritis surgery (average age 66 years), and they were grouped and numbered. All samples were confirmed by pathological examination, and the control group was patients with meniscal injury and cruciate ligament injury who underwent arthroscopic surgery. All sample numbers were stored in a -80°C low-temperature refrigerator.

[0059] 2. Expression situation analysis method

[0060] 2.1 Extract the total RNA from the tissue sample, the same as the extraction method in Example 1.

[0061] 2.2 Synthesis of cDNA by reverse transcription

[0062] use III Reverse Transcriptase (invitrogen, Cat. No. 18080-044) was used for cDNA reverse transcription, and the experimental operation was carried out accordi...

Embodiment 3

[0077] Embodiment 3 osteoarthritis diagnostic kit

[0078] Based on the primer set obtained in Example 2, the kit for diagnosing osteoarthritis according to the present invention was assembled, and the kit included the primer set for specifically amplifying hsa_circ_0045714 and hsa_circ_0005567 as shown in Table 1, specifically:

[0079] 1. The kit includes specific amplification hsa_circ_0045714: SEQ ID NO.3 and SEQ ID NO.4;

[0080] 2. The kit includes specific amplification hsa_circ_0005567: SEQ ID NO.5 and SEQ ID NO.6;

[0081] 3. The kit includes specific amplification hsa_circ_0045714 and hsa_circ_0005567: SEQ ID NO.3, SEQ ID NO.4 and SEQ ID NO.5, SEQ ID NO.6.

[0082] The primer pair for specifically amplifying the housekeeping gene (GAPDH) is shown in SEQ ID NO: 7 and SEQ ID NO: 8; it also includes a SYBR Green polymerase chain reaction system, such as PCR buffer, SYBR Green fluorescent dye, and dNTPs. The composition of described PCR buffer is 25mM KCl, 2.5mM MgCl ...

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Abstract

The present invention discloses the application of hsa_circ_0045714 and / or hsa_circ_0005567 as molecular markers in the preparation of products for diagnosing or treating osteoarthritis, and further confirms that both hsa_circ_0045714 and hsa_circ_0005567 are up-regulated in osteoarthritis tissues. The present invention further discloses that hsa_circ_0045714 and / or hsa_circ_0005567 are used to prepare a diagnostic kit for detecting osteoarthritis, and the kit includes primers and instructions for specifically amplifying osteoarthritis-related circRNA. Using hsa_circ_0045714 and / or hsa_circ_0005567 to detect osteoarthritis can not only quickly and effectively achieve early detection, but also provide therapeutic targets and important basis for clinical applications such as gene therapy and drug therapy.

Description

technical field [0001] The present invention relates to the field of biomedicine, in particular to the application of hsa_circ_0045714 and / or hsa_circ_0005567 in the preparation of products for diagnosing or treating osteoarthritis. 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 h...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/158C12Q2600/178
Inventor 肖刻翁习生边焱焱冯宾
Owner PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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