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Exosomal biomarker for auxiliary diagnosis of SCA3/MJD and screening and identification method thereof

A technology of secretobiology and auxiliary diagnosis, which is applied in the direction of biochemical equipment and methods, microbial measurement/testing, DNA/RNA fragments, etc. Ineffective treatment and other issues

Active Publication Date: 2019-01-25
XIANGYA HOSPITAL CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because its pathogenesis, clinical classification, disease progression and other related mechanisms have not been fully elucidated, there is no specific treatment, and the mortality and disability rates are high, which has brought a heavy burden to patients, families and society.

Method used

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  • Exosomal biomarker for auxiliary diagnosis of SCA3/MJD and screening and identification method thereof
  • Exosomal biomarker for auxiliary diagnosis of SCA3/MJD and screening and identification method thereof
  • Exosomal biomarker for auxiliary diagnosis of SCA3/MJD and screening and identification method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] In Example 1, the differentially expressed miRNAs related to SCA3 / MJD were screened, and the specific operation process was as follows:

[0034] 1. Sample collection: plasma samples from 12 patients with SCA3 / MJD (including 3 subtypes: 3 cases of type 1, 6 cases of type 2, and 3 cases of type 3) and plasma samples of 12 age- and sex-matched healthy individuals Serve as control group; CSF samples from 12 SCA3 / MJD patients (including 3 subtypes: 3 Type 1, 6 Type 2, 3 Type 3) and 10 age- and sex-matched aneurysm / migraine patients The cerebrospinal fluid samples were used as the control group.

[0035] Plasma samples: Collect 10ml of whole blood from SCA3 / MJD patients and control groups with anticoagulant tubes, centrifuge at 3000rpm at 4°C for 15min, separate the plasma for later use, and complete within 2 hours.

[0036] Cerebrospinal fluid samples: Collect 10-15ml of cerebrospinal fluid from SCA3 / MJD patients and the control group with sterile centrifuge tubes, and comp...

Embodiment 2

[0060] This example uses qPCR to verify differentially expressed miRNAs, wherein the verified miRNAs include novel-mir-9034, novel-mir-7660, novel-mir-5219, novel-mir-8421, novel-mir-769, novel-mir -7014, novel-mir-6628, novel-mir-4682, novel-mir-2738, novel-mir-1643, has-miR-92b-3p, has-miR-486-3p, has-miR-484, has -miR-378g, has-miR-375, has-miR-328-3p, has-miR-3074-5p, has-miR-24-3p, has-miR-206, has-miR-204-5p, has -miR-151a-3p, has-miR-146b-5p, has-miR-142-3p, has-miR-1290, has-miR-1268a, has-miR-122-5p, has-let-7d-3p , the specific operation process of qPCR verification is as follows:

[0061] 1. Sample collection: 18 plasma samples of SCA3 / MJD patients and 16 plasma samples of age- and sex-matched healthy people were used as control group; 10ml of whole blood was collected from SCA3 / MJD patients and control group with anticoagulant tube, 3000rpm 4 Centrifuge at ℃ for 15 minutes, separate the plasma for later use, and complete within 2 hours.

[0062] 2. The extractio...

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Abstract

The invention discloses an exocrine biomarker for auxiliary diagnosis of SCA3 / MJD and a screening and identification method thereof, belonging to the field of cell and molecular biotechnology. The total RNA is extracted from the plasma and cerebrospinal fluid exosomes of SCA3 / MJD patient, the differentially expressed miRNAs were screened and identified by Small RNA Sequencing (sRNA seq) and verified by real-time fluorescent real-time PCR (qPCR). It confirms that the novel-mir-7014 can be use as a potential biomarker to guide that clinical classification of SCA3 / MJD and to reflect the severityof the disease. It will provide a new specific auxiliary diagnostic method for SCA3 / MJD, and has potential clinical application value in condition monitor and adjuvant therapy.

Description

technical field [0001] The present invention relates to the field of cell and molecular biology technology, in particular to an exosome biomarker for auxiliary diagnosis of SCA3 / MJD and a screening and identification method thereof. Background technique [0002] Hereditary spinocerebellar ataxias (spinocerebellar ataxias, SCAs) is a common neurodegenerative disease, especially spinocerebellar ataxia type 3 / Machado-Joseph disease (spinocerebellar ataxia type 3 / Machado-Joseph disease, SCA3 / MJD) is the most common type, which accounts for almost 60-70% of SCAs patients in the Chinese population. Some scholars have divided SCA3 / MJD into the following five subtypes: type 1: usually onset before the age of 20, progresses rapidly, and is characterized by pyramidal tract signs (myotonia and muscle spasm) and extrapyramidal features (bradykinesia and muscle tension). disorder) combined with ataxia as the main manifestation; type 2 usually develops between the ages of 20 and 50, with...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12N15/113
CPCC12Q1/6883C12Q2600/158C12Q2600/178
Inventor 江泓侯晓灿陈召裘嵘唐北沙
Owner XIANGYA HOSPITAL CENT SOUTH UNIV
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