Establishment of OA marker

A marker and marker technology, applied in the field of gene transcriptomics and proteomics, can solve the problems of inability to display deep cartilage and subchondral bone, and clinical application limitations.

Inactive Publication Date: 2018-04-13
南宁科城汇信息科技有限公司
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  • Abstract
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Problems solved by technology

However, its clinical application is also limited due to the invasiveness of the exa

Method used

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Embodiment

[0029] The establishment of such OA markers;

[0030] Consists of the following steps:

[0031] 1. Application of iTRAQ labeling combined with MALDI-TOF MS / MS technology in STR / ort model of osteoarthritis

[0032] ①Sample preparation: 10 STR / ort model mice and 10 control mice each were raised in a semi-barrier system according to the feeding requirements of clean-grade animals; raised to 20 weeks old (adult), blood was collected from each mouse at 0.9- 1.5 mL; After the blood sample was allowed to stand at 4°C for 1 hour, centrifuge at 5000 r / min for 5 minutes, take the supernatant, put 50 μL / tube into a 0.2 mL EP tube, mark it well, and store it at -80°C for later use;

[0033] ②Extract the serum of the STR / ort model group and the control group, mix the two groups in equal volumes, and use the acetone precipitation method to precipitate and quantify the protein, so that each group contains about 100ug of total protein, and trypsinize and reduce the alkyl group in turn Chemi...

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Abstract

The invention discloses establishment of an OA marker. The establishment of the OA marker includes: first, application of an iTRAQ label and an MALDI-TOF MS/MS technology in an osteoarthritis STR/ortmodel: (1) preparation of a sample, (2) extraction of serum of an STR/ort model set and serum of a control group, (3) identification of differential protein of the serum of the STR/ort model set and differential protein of the serum of the control group according to difference; second, verification of the OA marker: (1) acquiring of a sample of a patient, (2) use of Western blot, and (3) one-by-one detection of an OA candidate marker by use of Western blot; and third, confirmation of the OA marker: (1) collection of the sample of the patient, (2) detection by use of an ELISA method, and (3) establishment of the OA marker. The marker is confirmed by population samples, value of these OA candidate markers on diagnosis of OA is evaluated, clinical dependency of the OA candidate markers is analyzed, and feasibility of the OA candidate markers in clinical application of prevention of OA, early diagnosis and the like and significance of the OA candidate markers in early warning of health ofbone joints of population are proposed.

Description

technical field [0001] The invention relates to the fields of gene transcriptomics and proteomics, especially the establishment of an OA marker. Background technique [0002] Osteoarthritis (OA), also known as degenerative bone joint disease, is one of the most common joint diseases, mainly manifested as slowly developing pain, stiffness, swelling of the joints, accompanied by joint dysfunction, and even disability[1- 2]. The pathogenesis of OA is that joints are affected by genetic, biological and mechanical events (such as genetics, metabolic abnormalities, strain, trauma, etc.), which induce changes in cells and matrix in terms of morphology, biomechanics, and molecules. It leads to an imbalance in the degradation and synthesis of articular chondrocytes, extracellular matrix and subchondral bone. The outcome is softening, fibrosis, ulceration and loss of chondrocytes. After losing the protective effect of cartilage, the subchondral bone hardens and becomes ivory, eventu...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/543
CPCG01N33/6893G01N33/543G01N2560/00G01N2570/00G01N2800/105
Inventor 于思创王海云
Owner 南宁科城汇信息科技有限公司
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