Circulating microRNA biomarker for subarachnoid hemorrhage and application thereof

A technology of subarachnoid space and mir-4532, applied in biochemical equipment and methods, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of impaired quality of life and inability to return

Active Publication Date: 2018-11-02
THE CHINESE UNIVERSITY OF HONG KONG +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

More than a third of SAH patients experience cognitive impairment, impaired quality of life, and may not be able to return to pre-illness jobs

Method used

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  • Circulating microRNA biomarker for subarachnoid hemorrhage and application thereof
  • Circulating microRNA biomarker for subarachnoid hemorrhage and application thereof
  • Circulating microRNA biomarker for subarachnoid hemorrhage and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Embodiment 1 patient and sample

[0037] The study was approved by the Joint Committee on Clinical Research Ethics of the New Territories East Hospital Cluster-Chinese University of Hong Kong (NTEC-CUHK), and written informed consent was obtained from all participating patients or their closest blood relatives. On day 7 after SAH in healthy controls (N=20), subarachnoid hemorrhage (SAH) patients with delayed cerebral infarction (DCI) (N=20), SAH patients without DCI (N=20 ) on the 7th day after SAH, peripheral blood miRNA was collected. SAH patients were from Prince of Wales Hospital, Chinese University of Hong Kong from 2012 to 2013. Ruptured brains were diagnosed by computed tomography angiography (CTA). In contrast to SAH patients, healthy controls (n=20) were recruited from family members of SAH patients without significant medical risks, including no smoking history and hypertension. The condition and signs of the patients are shown in Table 1.

[0038] Table 1...

Embodiment 2

[0042] Embodiment 2 Delayed cerebral infarction (DCI) definition

[0043] In this study, delayed cerebral infarction (DCI) was defined as cerebral infarction identified by CT after excluding procedure related infarction (Wong GK et al., Journal of neurology, neurosurgery, and psychiatry. 2012; 83 : 1112-1117; Vergouwen MD et al., Proposal of multidisciplinary research group. Stroke; a journal of cerebral circulation. 2010; 41:2391-2395). Surgery-related infarction refers to the hypodense area shown on postoperative CT at about 12-24 hours after treatment. All patients participating in the study underwent CT time-lapse scanning after 2-3 weeks. DCI was diagnosed by consensus of two neuroradiologists.

Embodiment 3

[0044] Embodiment 3 real-time fluorescent quantitative PCR

[0045] Peripheral blood samples were obtained using EDTA tubes according to standard procedures. Samples were immediately placed on ice and centrifuged at 1000 g for 15 minutes at 4°C. Plasma fractions were collected and stored in aliquots at -80°C. RNA isolation and quantitative real-time PCR were performed according to conventional methods, including by adding 20% ​​w / v CaCl 2 (Sigma, St. Louis, MO) was added to plasma samples to prepare serum, then clotted overnight and centrifuged. Total RNA was isolated from the resulting serum samples using the miRNeasy serum / plasma kit (Qiagen, Valencia, CA) according to the manufacturer's instructions. The amount of extracted RNA was measured by Nanodrop 2000 UV-Vis spectrophotometer (Thermo Scientific, Waltham, MA). Ct values ​​and melting curves were analyzed using Applied Biosystems SDS software (Applied Biosystems, Waltham, MA).

[0046] A total of 28 specific miRNAs...

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Abstract

The invention provides a biomarker of subarachnoid hemorrhage indicating poor prognosis. The poor prognosis is delayed cerebral infarction, and the marker consists of the following miRNAs: miR-4463, miR-4532 and miR-1290, and may also comprises miR-4793. The invention also provides a method for diagnosing subarachnoid hemorrhage or prognosis thereof by using the biomarker. The invention also provides a kit and a chip containing the biomarker for diagnosing subarachnoid hemorrhage or prognosis thereof, and a preparation method thereof.

Description

technical field [0001] The invention relates to the fields of molecular biology and preparation of disease diagnosis reagents. Specifically, the present invention relates to microRNA biomarkers of subarachnoid hemorrhage and subsequent delayed cerebral infarction, and the preparation of said microRNA for diagnosing or detecting subarachnoid hemorrhage and subsequent delayed cerebral infarction The application of the infarction kit or gene chip is especially used in peripheral blood samples, including serum and plasma samples. Background technique [0002] Subarachnoid hemorrhage (SAH) accounts for about 3-5% of strokes, and is an important cause of stroke in young people, bringing serious social and economic burden worldwide. More than one-third of people with SAH experience cognitive impairment, impaired quality of life, and may not be able to return to their pre-illness jobs. [0003] Delayed cerebral infarction (DCI) occurs in up to 44% of SAH patients and usually begin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12N15/113
CPCC12Q1/6883C12Q2600/118C12Q2600/158C12Q2600/172A61K31/7088C12N15/113C12Q1/68
Inventor 黄国柱马金龙潘伟生陈伟仪路钢熊志强
Owner THE CHINESE UNIVERSITY OF HONG KONG
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