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Urine exosome hsa-micro RNA486-3p and application thereof

A technology of exosomes and urine, applied in the field of medicine, can solve the problems of high price, impossibility of popularization, and long time-consuming, and achieve the effect of low price, easy to carry and popularize

Inactive Publication Date: 2017-09-22
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-microRNA486-3p

Method used

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  • Urine exosome hsa-micro RNA486-3p and application thereof
  • Urine exosome hsa-micro RNA486-3p and application thereof
  • Urine exosome hsa-micro RNA486-3p and application thereof

Examples

Experimental program
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Embodiment 1

[0055] (1) Screening of candidate markers of urinary exosomes

[0056] 1. Acquisition and identification of urine exosomes

[0057] a. Obtaining method:

[0058] 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.

[0059] 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:

[0060] (1) Take the urine sample out from -80°C and put it on ice for later use;

[0061] (2) Centrifuge at room temperature for 20min (14,000rpm) to remove residual cells and debris;

[0062] (3) Transfer the supernatant to a new tube, add 1 / 3 volume of RiboTM exosome...

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Abstract

The invention discloses urine exosome hsa-micro RNA486-3p and application thereof, belonging to the technical field of medicines. The urine exosome hsa-micro RNA486-3p is based on an exosome liquid biopsy technology, utilizes a next generation sequencing technique of an Illumina Hiseq2500 sequencing platform, and is used for detecting differentially expressed micro RNA(s) of steroid-induced necrosis of femoral head in urine exosomes; the differentially expressed micro RNA(s) can be verified by qRT-polymerase chain reaction (PCR), so that a potential marker with obvious differential expression can be obtained; the sequence of the urine exosome hsa-micro RNA486-3p is as shown in SEQ ID No: 1. The urine exosomes are extracted, and the change of the hsa-micro RNA486-3p in the urine exosomes is analyzed by using the next generation sequencing technique, so that the steroid-induced necrosis of femoral head can be found out earlier than magnetic resonance imaging (MRI); the urine exosome hsa-micro RNA486-3p is low in price and convenient to carry and popularize.

Description

technical field [0001] The invention belongs to the technical field of medicine, and relates to a urine exosome hsa-microRNA486-3p, a biomarker for ultra-early diagnosis of steroid-induced necrosis of the femoral head, in particular to a urine exosome hsa-microRNA486-3p and its application . 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): C12Q1/68C12N15/113
CPCC12Q1/6883C12Q2600/158
Inventor 陈鹏何伟陈德龙陈镇秋王海彬姜山邹许亭
Owner THE FIRST AFFILIATED HOSPITAL OF GUANGZHOU UNIV OF CHINESE MEDICINE