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An exosome biomarker for auxiliary diagnosis of sca3/mjd and its screening and identification method
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An exosome biological, auxiliary diagnosis technology, applied in biochemical equipment and methods, microbial determination/examination, DNA/RNA fragments, etc. Ineffective treatment, etc.
Active Publication Date: 2021-06-29
XIANGYA HOSPITAL CENT SOUTH UNIV
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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.
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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 Served 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.
[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 exosome biomarker for auxiliary diagnosis of SCA3 / MJD and a screening and identification method thereof, belonging to the technical field of cell and molecular biology. The present invention extracts total RNA from plasma and cerebrospinal fluid exosomes of SCA3 / MJD patients, applies Small RNA Sequencing (sRNA seq) to screen and identify differentially expressed miRNAs, and combines real-time fluorescence quantitative PCR (Quantitative Real-time PCR, qPCR ) verification, confirmed that novel‑mir‑7014 can be used as a potential biomarker to guide the clinical classification of SCA3 / MJD and reflect the severity of the disease, and will provide a new specific auxiliary diagnostic method for SCA3 / MJD. and adjuvant therapy have potential clinical application value.
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...
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