A kind of ischemic retinopathy detection biomarker, detection kit and application

A technology for retinopathy and ischemia, applied in the field of biomedical detection, which can solve the problems such as the lack of circRNA-Wdr37

Active Publication Date: 2021-04-23
南京医科大学眼科医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, there is no report of circRNA-Wdr37 as a biomarker for the diagnosis of ischemic retinopathy

Method used

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  • A kind of ischemic retinopathy detection biomarker, detection kit and application
  • A kind of ischemic retinopathy detection biomarker, detection kit and application
  • A kind of ischemic retinopathy detection biomarker, detection kit and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Verification of the correlation between circRNA-Wdr37 and ischemic retinopathy

[0037] CircRNA microarray analysis screens and verifies circRNAs related to ischemic retinopathy.

[0038] The first step: sample preparation: the mouse retinal ischemia-reperfusion (IR) animal model was constructed, and the retinal tissues of the experimental group (IR, n=3) and the control group (Ctrl, n=3) were collected. RNA was extracted with TRIzol (Invitrogen) reagent and stored at -80°C for future use.

[0039] Step 2: Differentially expressed circRNA screening:

[0040] Using the circRNA expression profile chip of Aglient Company in the United States, the circRNA related to the occurrence of ischemic retinopathy was analyzed; the specific steps of the analysis were: using a labeling enzyme to label the circRNA with a fluorescent group to obtain a fluorescent probe for hybridization with the chip. Use the MAUI hybridization instrument and chip hybridization; use the Gene...

Embodiment 2

[0045] Example 2 Detection of expression of circRNA-Wdr37 in blood cells and plasma

[0046] Step 1: Separation of serum and plasma samples

[0047] Blood samples of 200 cases of each patient in the experimental group (primary open-angle glaucoma, central vein occlusion, and diabetic retinopathy) and the control group (cataract) were collected, and serum and serum were separated by centrifugation using heparin anticoagulation tubes. Plasma RNA for detection of circRNA. The centrifugation conditions were 4°C, 12,000 rpm, 10 min.

[0048] Step 2: RNA Extraction from Serum and Plasma Samples

[0049] Add TRIzol to the separated blood cells and plasma samples respectively, and place at room temperature for 10 minutes to fully lyse the samples (Note: If the next step is not performed, the samples can be stored at -70°C for long-term storage). Add 200 μl of chloroform to every 1ml of TRIzol, vibrate vigorously and mix well, then place at room temperature for 3-5min to allow natur...

Embodiment 3

[0075] Example 3 Feasible application of circRNA-Wdr37 as a marker for prognosis assessment

[0076] The first step; obtain the blood sample to be tested

[0077] Sera from 200 patients with each disease were collected before and after treatment for ischemic retinopathy (primary open-angle glaucoma, central vein occlusion, and diabetic retinopathy).

[0078] Step 2: Extract RNA from the blood sample to be tested

[0079] a) Serum sample or frozen serum Take 0.25ml sample and transfer it to a centrifuge tube, add 1ml Trizol, pipette up and down repeatedly until the cells are completely lysed.

[0080] b) Add chloroform (sample solution + 1 / 5 volume of Trizol Reagent volume) to cover the centrifuge tube tightly, vibrate vigorously for 15 sec, and let stand at room temperature for 5 min;

[0081] c) Centrifuge at 4°C, 12,000g×15min. Carefully take out the centrifuge tube from the centrifuge. At this time, the homogenate is divided into three layers, namely: a colorless supernat...

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Abstract

The invention belongs to the technical field of biomedical detection, and in particular relates to a biomarker for detection of ischemic retinopathy, a detection kit and its application. The invention discloses the application of circRNA-Wdr37 in preparing a diagnostic reagent for ischemic retinopathy and a diagnostic kit for ischemic retinopathy. Through high-throughput sequencing and quantitative PCR verification, the present invention finds that circRNA‑Wdr37 is significantly correlated with the occurrence of ischemic retinopathy. The present invention uses real-time fluorescent quantitative PCR technology to detect the relative content of circRNA-Wdr37 in aqueous humor, and can be used for the screening of patients with clinical ischemic retinopathy, and provides a theoretical basis for early intervention of ischemic retinopathy.

Description

technical field [0001] The invention belongs to the technical field of biomedical detection, and in particular relates to a detection biomarker for ischemic retinopathy, a detection kit and its application, which can be used to diagnose and develop human ischemic retinopathy according to the expression level of individual circRNA-Wdr37 judgment. Background technique [0002] Ischemic retinopathy is a common ophthalmic disease mainly composed of central retinal vein occlusion (CRVO), branch vein occlusion (BRVO), ocular hypertension glaucoma, diabetic retinopathy (DR), ocular ischemic syndrome and aortic Inflammatory fundus lesions and other retinal vascular poor perfusion lesions caused. After the occurrence of retinal ischemia, the ischemia-hypoxic damage leads to the damage of retinal ganglion cells and other cell functions, and the decline of visual function, which can seriously affect the quality of life of patients. In the early stage of chronic ischemic retinopathy, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883C12Q1/6851C12N15/113A61K31/7088A61P27/02
CPCA61K31/7088A61P27/02C12Q1/6851C12Q1/6883C12Q2600/118C12Q2600/158C12Q2600/178C12Q2531/113C12Q2563/107C12Q2521/107
Inventor 蒋沁颜标姚牧笛张秋阳马严朱妍张荟颖
Owner 南京医科大学眼科医院
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