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Application of interferon stimulating gene 15 in preparation of products for early diagnosis of autoimmune eye diseases

An autoimmune and interferon-based technology, applied in disease diagnosis, material testing products, biochemical equipment and methods, etc., can solve the problems of poor economic burden, poor quality of life, no gold standard, missed diagnosis and misdiagnosis, etc., to reduce the misdiagnosis rate and Blindness rates, effects of sensitive clinical strategies

Inactive Publication Date: 2022-03-25
ZHONGSHAN OPHTHALMIC CENT SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the diagnosis of autoimmune eye diseases is very complicated and there is no gold standard. It is very dependent on the diagnosis and treatment level of clinical ophthalmologists, and misdiagnosis and misdiagnosis are very easy to occur. The misdiagnosis rate of literature reports is as high as 85%, which has caused a great impact on the prognosis and vision of patients. damage, resulting in severe economic burden and poor quality of life

Method used

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  • Application of interferon stimulating gene 15 in preparation of products for early diagnosis of autoimmune eye diseases
  • Application of interferon stimulating gene 15 in preparation of products for early diagnosis of autoimmune eye diseases
  • Application of interferon stimulating gene 15 in preparation of products for early diagnosis of autoimmune eye diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 Detection of differential expression of interferon-stimulated gene 15 by single-cell sequencing technology based on microfluidic system

[0040] Using the single-cell sequencing technology based on the microfluidic system, perform 10X single-cell sequencing analysis on each sample to obtain differential genes. The main steps include:

[0041] (1) Cell separation and capture: 10X Genomics Chromium Controller is used for droplet generation, and single cells are wrapped into droplets to form water-in-oil single-cell droplets; the specific process is: Gel Beads with Barcode are passed through a process The sample port enters the microfluidic system at a constant speed, and the cells and enzymes to be sorted enter the microfluidic system at a certain time interval through another sample port, and combine with Gel Beads, and then enter the oil phase to form GEMs (Gel Bead in emulsion) water-in-oil droplet structure;

[0042] (2) Microfluidic chip amplification: aft...

Embodiment 2

[0046] Example 2 Flow cytometry analysis of differential expression of interferon-stimulated gene 15 protein

[0047] Perform intracellular staining flow cytometry analysis on each sample to verify the expression of interferon-stimulated gene 15 protein, the main steps include:

[0048] (1) pair 1×10 6 Resuspend each PBMC with 200uL PBS, add 2μl each of CD3-FITC, CD19-FITC, CD56-FITC, CD14-APC-H7, CD16-AF647 monoclonal antibodies for surface marker staining, and incubate on ice for 30 minutes in the dark;

[0049] (2) washing the cells 3 times;

[0050] (3) Purchase the cell membrane disruption kit from BD biosciences, resuspend the cells with 100 μl Cytofix / Cytoperm fixation / permeation solution, and incubate on ice in the dark for 20 minutes;

[0051] (4) Wash the cells 3 times with Wash / Perm buffer;

[0052] (5) Resuspend the cells with 200 μl buffer, add 10 μl interferon-stimulated gene 15 protein-PE antibody for intracellular staining, and incubate on ice for 30 minutes...

Embodiment 3

[0056] Example 3 ELISA detection and analysis of differential expression of interferon-stimulated gene 15 protein

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Abstract

The invention discloses application of an interferon stimulating gene 15 in preparation of a product for early diagnosis of autoimmune eye diseases. The interferon-stimulated gene 15 or the expression product thereof is used as the marker related to the autoimmune eye disease, whether a subject suffers from the autoimmune eye disease or not can be judged in the early stage, effective treatment can be carried out in advance, the misdiagnosis rate and blindness rate of the autoimmune eye disease patient are reduced, and the autoimmune eye disease treatment effect is improved. And a more convenient, rapid and sensitive clinical strategy can be provided.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the application of interferon-stimulated gene 15 in the preparation of early diagnosis products for autoimmune eye diseases. Background technique [0002] Human Interferon Stimulated Gene 15 (Interferon Stimulated Gene 15) is a ubiquitin-like protein that can be covalently conjugated to the target protein by protease cleavage to expose the carboxy-terminal sequence in the cell, and carry out ISG modification on it; it can also be directly secreted Exercising immunomodulatory function extracellularly. At present, there is no report on the use of interferon-stimulated gene 15 or its expression product as a diagnostic molecule for autoimmune eye diseases. [0003] Autoimmune eye disease is a kind of vision-threatening intraocular inflammation, which accounts for about 10% of global blindness. It is caused by various reasons such as autoimmune imbalance. It is difficult to t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883G01N33/68
CPCC12Q1/6883G01N33/6893C12Q2600/158G01N2800/16G01N2800/24G01N2333/4706
Inventor 迟玮胡译心
Owner ZHONGSHAN OPHTHALMIC CENT SUN YAT SEN UNIV
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