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Hsa-microRNA06 of urinary exosome and application thereof

A technology for exosomes and urine, applied in the field of medicine, can solve the problems of high price, inability to popularize, and disease, and achieve the effects of low price, easy portability and promotion

Inactive Publication Date: 2018-11-20
THE FIRST AFFILIATED HOSPITAL OF GUANGZHOU UNIV OF CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0039] In order to overcome the shortcomings and deficiencies in the prior art: 1. The disease has already occurred at the time of diagnosis; 2. It is expensive and takes a long time; 3. The MRI equipment itself is expensive and cannot be popularized; the purpose of the present invention is to provide a urine exosome hsa-microRNA206

Method used

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  • Hsa-microRNA06 of urinary exosome and application thereof
  • Hsa-microRNA06 of urinary exosome and application thereof
  • Hsa-microRNA06 of urinary exosome and application thereof

Examples

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Embodiment 1

[0056] (1) Screening of candidate markers of urinary exosomes

[0057] 1. Acquisition and identification of urine exosomes

[0058] a. Obtaining method:

[0059] Urine was collected from healthy people, no femoral head necrosis after steroid use, and steroid-induced femoral head necrosis (ARCO IV stage) requiring total hip arthroplasty, and then exosomes isolation kit was used to extract the exosomes in the urine.

[0060] Use the exoRNeasy Serum / Plasma Kits Exosome Isolation Kit from Qiagen (Cat. No.: 77064) or the RiboTM Exosome Isolation Kit from Guangzhou Ruibo Biotechnology Co., Ltd. (Cat. No.: C10110), and refer to the kit instructions for specific steps . The steps are briefly described as follows:

[0061] (1) Take the urine sample out from -80°C and put it on ice for later use;

[0062] (2) Centrifuge at room temperature for 20min (14,000rpm) to remove residual cells and debris;

[0063] (3) Transfer the supernatant to a new tube, add 1 / 3 volume of RiboTM exosome...

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Abstract

The invention discloses hsa-microRNA06 of a urinary exosome and the application of the hsa-microRNA06, and belongs to the technical field of medicines. The invention makes use of a next generation ofsequencing technique of an Illumina Hiseq2500 sequencing platform to find out differentially expressed microRNA(s) in femoral head necrosis in the urinary exosome based on an exosome liquid biopsy technique; qRT-PCR verifies the differentially expressed microRNA(s) to obtain potential marks obvious in differential expression; the sequence of the hsa-miR06 of the urinary exosome is as shown in SEQID NO:1. The urinary exosome is extracted, and variations of the hsa-miR06 in the urinary exosome are analyzed by using the next generation of sequencing technique, so that the femoral head necrosis can be found earlier than MRI (Magnetic Resonance Imaging); the cost is low, and the carrying and promotion are convenient.

Description

technical field [0001] The invention belongs to the technical field of medicine, and relates to a urine exosome hsa-microRNA206 as a biomarker for ultra-early diagnosis of steroid-induced necrosis of the femoral head, in particular to a urine exosome hsa-microRNA206 and an application thereof. Background technique [0002] Osteonecrosis of the femoral head (ONFH) is an orthopedic disease with a high disability rate. It occurs frequently in young and middle-aged people aged 30-50. It is likely to cause the loss of working ability of patients and bring a heavy burden to the family and society[1,2]. If it is not detected in time and intervention measures are not taken, 80% of ONFH will rapidly progress to the late stage of osteonecrosis (arthritic stage), and finally receive total hip replacement treatment [3]. Therefore, it is an urgent problem to be solved in clinical work to detect necrosis of the femoral head as early as possible by using non-invasive detection methods, and...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12N15/113
CPCC12Q1/6883C12Q2600/158C12Q2600/178
Inventor 陈鹏陈德龙何伟王海彬姜山邹许亭
Owner THE FIRST AFFILIATED HOSPITAL OF GUANGZHOU UNIV OF CHINESE MEDICINE