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Application of brain cell-derived exosome circular RNA in serum as Alzheimer's disease diagnostic marker

A technology for Alzheimer's disease and exosomes, applied in the field of medical diagnosis, which can solve the problems of large patient damage, low utilization rate, and few product types

Active Publication Date: 2021-12-10
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

AD diagnostic technology is immature, and there are few types of products. Currently, mainstream products focus on β-amyloid and Tau protein markers in cerebrospinal fluid, but lumbar puncture is required for cerebrospinal fluid samples. lower

Method used

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  • Application of brain cell-derived exosome circular RNA in serum as Alzheimer's disease diagnostic marker
  • Application of brain cell-derived exosome circular RNA in serum as Alzheimer's disease diagnostic marker
  • Application of brain cell-derived exosome circular RNA in serum as Alzheimer's disease diagnostic marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Identification of brain-derived exosome circular RNA expression profile in serum

[0025] Collect 3 cases of Alzheimer's disease patients and 3 cases of non-Alzheimer's disease patients with clinical symptoms similar to Alzheimer's disease serum, each 5ml, separate exosomes, and use high-throughput sequencing to detect the circularity in exosomes RNA (circRNA) expression.

[0026] Step 1: Isolation of brain-derived exosomal RNA in serum:

[0027] 1) 5ml of serum from each test object was centrifuged at a speed of 300g and room temperature for 10 minutes to remove the precipitate and retain the supernatant;

[0028] 2) Centrifuge the supernatant obtained in the previous step for 30 minutes at a rotation speed of 10,000 g and room temperature, remove the precipitate and retain the supernatant;

[0029] 4) Add magnetic beads coupled with L1CAM (L1 cell adhesion molecule), GLAST (Glutamate aspartate transporter) and TMEM119 (transmembrane protein 119) antibodies...

Embodiment 2

[0035] Example 2 Quantitative detection of brain-derived exosomal circular RNA that can be used as an early diagnosis of Alzheimer's disease in serum

[0036] Collect 100 cases of Alzheimer's disease patients and 50 cases of non-Alzheimer's disease patients with clinical symptoms similar to Alzheimer's disease serum, each 5ml, separate exosomes from brain cells, and use high-throughput sequencing to detect exosomes. In vivo circular RNA (circRNA) expression.

[0037] Step 1: Isolation of brain-derived exosomal RNA in serum:

[0038] 1) 5ml of serum from each test object was centrifuged at a speed of 300g and room temperature for 10 minutes to remove the precipitate and retain the supernatant;

[0039] 2) Centrifuge the supernatant obtained in the previous step for 30 minutes at a rotation speed of 10,000 g and room temperature, remove the precipitate and retain the supernatant;

[0040]4) Add magnetic beads coupled with L1CAM (L1 cell adhesion molecule), GLAST (Glutamate asp...

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Abstract

The invention discloses application of brain cell-derived exosome circular RNA in serum as an Alzheimer's disease diagnostic marker. The invention relates to application of a reagent for detecting any one or more of hsa_circ_0029426 and hsa_circ_0044837 in preparing an Alzheimer's disease auxiliary diagnostic reagent. High-throughput sequencing and quantitative PCR analysis in nearly 100 patients finds that two kinds of circular RNA in exosome in serum can effectively distinguish Alzheimer's disease patients from normal people and non-Alzheimer's disease patients with clinical symptoms similar to Alzheimer's disease, and the area under an ROC curve is greater than 0.90. Therefore, the two kinds of circular RNA or a combination thereof can be used as detection markers for Alzheimer's disease diagnosis.

Description

technical field [0001] The invention belongs to the field of medical diagnosis, and relates to a circular RNA marker in exosomes related to Alzheimer's disease and its application. Background technique [0002] Alzheimer's disease (AD) is a neurodegenerative disease commonly seen in the elderly. Cognitive impairment and memory impairment are the main clinical symptoms. The prevalence of Alzheimer's disease is age-related, with an increase of 19% in the elderly population after the age of 85. Affected by the aging of the population, the number of patients with Alzheimer's disease continues to increase. AD diagnostic technology is immature, and there are few types of products. The current mainstream products are concentrated in β-amyloid and Tau protein markers in cerebrospinal fluid. However, lumbar puncture is required for cerebrospinal fluid samples, which is more harmful to patients. lower. [0003] Exosomes are tiny vesicles with a lipid bilayer membrane structure and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/11C12Q1/6883C12Q1/6851
CPCC12N15/113C12Q1/6883C12Q1/6851C12N2310/531C12Q2600/158C12Q2600/178C12Q2525/307C12Q2531/113Y02A50/30
Inventor 梁宏伟渠爽卢庚甘文华王冯娟
Owner CHINA PHARM UNIV
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