Cerebral stroke LncRNA biomarker and application thereof

A marker and stroke technology, applied in DNA/RNA fragments, recombinant DNA technology, microbial determination/inspection, etc.

Active Publication Date: 2019-01-04
JIANGSU UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the research on lncRNA markers in the regulation mechanism of stroke is still in its infancy

Method used

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  • Cerebral stroke LncRNA biomarker and application thereof
  • Cerebral stroke LncRNA biomarker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] 1. sample collection

[0022] The blood of 100 cases of healthy people and stroke patients were collected, and the patients gave informed consent, and all the above samples were obtained with the consent of the organizational ethics committee.

[0023] 2. RNA sample preparation

[0024] (1) After clinical serum samples are collected, centrifuge at 12,000 rpm for 10 minutes at high speed, repeat twice, and store the obtained serum samples at -80°C;

[0025] (2) Thaw frozen serum samples at 4°C;

[0026] (3) Pipette 250µL serum sample to 1.5m1EP tube, add 750µL Trizol LS Reagent, mix by pipetting, and let stand for 5min;

[0027] (4) Add chloroform (CHCl3: Trizol 250µL:750µL), invert and mix well, and let stand for 15min;

[0028] (5) 4°C, 12000rpm, centrifuge for 15min;

[0029] (6) Carefully draw the upper liquid into a new EP tube;

[0030] (7) Add an appropriate amount of isopropanol (isopropanol: trizo1500 µL: 750 µL), invert and mix well, and let stand for 10...

Embodiment 2

[0051] Example 2: Expression of marker LncRNA in stroke tissue

[0052] 1. Sample Collection

[0053] According to the method of Example 1, 50 cases of blood from healthy people and blood from stroke patients were collected

[0054] 2. RNA extraction steps are as described in Example 1.

[0055] 3. The steps of reverse transcription are as described in Example 1.

[0056] 4. PCR amplification

[0057] Prepare PCR reaction system: 12.5 µL of 2×qPCR mixture, 2.0 µL of gene primers, 2.5 µL of reverse transcription products, ddH 2 08.0 µL.

[0058] The PCR reaction conditions were as follows: pre-denaturation at 95°C for 10 min, (denaturation at 95°C for 15 s, annealing at 60°C for 60 s) x 40 cycles, extension at 60°C for 5 min. Using SYBR Green as a fluorescent marker, the PCR reaction was carried out on a Light Cycler fluorescence quantitative PCR instrument, the target band was determined by melting curve analysis and electrophoresis, and the relative quantification was ...

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Abstract

The invention relates to the field of biological detection, in particular to biological molecular detection and more particularly relates to a cerebral stroke LncRNA biomarker and application thereof.A specific amplification primer pair of the LncRNA biomarker is high in expression in blood of cerebral stroke patients. The biomarker has a significant application prospect in preparation of products for cerebral stroke auxiliary assessment or cerebral stroke early assessment.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to biological molecular detection, and more specifically to a stroke LncRNA biomarker and its application. Background technique [0002] Cerebral stroke is an acute cerebrovascular disease, which is a group of diseases that cause brain tissue damage due to sudden rupture of blood vessels in the brain or blockage of blood vessels that prevent blood from flowing into the brain, including ischemic and hemorrhagic strokes. The incidence of ischemic stroke is higher than that of hemorrhagic stroke, accounting for 60% to 70% of the total number of strokes. Occlusion and stenosis of the internal carotid artery and vertebral artery can cause ischemic stroke, most of which are over 40 years old, more men than women, and severe cases can cause death. Mortality from hemorrhagic stroke is high. According to the survey, stroke has become the number one cause of death in my country combined ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12N15/113
CPCC12Q1/6883C12Q2600/158C12Q2600/178
Inventor 申红星邵世和娄培安刘庭君肖卉
Owner JIANGSU UNIV
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