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Application of lncRNA (long non-coding ribonucleic acid) to diagnosing cerebral ischemic stroke

An ischemic stroke, specific technology, applied in the direction of DNA / RNA fragments, recombinant DNA technology, microbial measurement / inspection, etc., can solve the problem of a large number of lncRNAs, and achieve the effect of improving accuracy

Active Publication Date: 2017-01-11
THE FIRST AFFILIATED HOSPITAL OF SHANTOU UNIV MEDICAL COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] A number of studies have confirmed that certain 1ncRNAs play an important role in human ischemic stroke, which is mainly reflected in the close relationship between various genes and signaling molecules involved in the regulation of angiogenesis. Research in disease is still in its infancy

Method used

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  • Application of lncRNA (long non-coding ribonucleic acid) to diagnosing cerebral ischemic stroke
  • Application of lncRNA (long non-coding ribonucleic acid) to diagnosing cerebral ischemic stroke

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Screening for Gene Markers Related to Ischemic Stroke

[0037] 1. Sample collection

[0038] 10 pairs of normal human blood and blood of ischemic stroke patients were collected, and the patients all gave informed consent, and all the above samples were obtained with the consent of the organizational ethics committee.

[0039] 2. Preparation of RNA samples

[0040] 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;

[0041] 2) Thaw frozen serum samples at 4°C;

[0042] 3) Pipette 250 μl serum sample to 1.5 ml EP tube, add 750 μl Trizol LS Reagent, mix by pipetting, and let stand for 5 minutes;

[0043] 4) Add chloroform (CHCl 3 : trizol 250μ1:750μ1), mix upside down, let stand for 15min;

[0044] 5) 4°C, 12000rpm, centrifuge for 15min;

[0045] 6) Carefully draw the upper liquid into a new EP tube;

[0046] 7) Add appropriate amount of isopropa...

Embodiment 2

[0061] Example 2 QPCR sequencing to verify the differential expression of the LOC101929707 gene

[0062] 1. Large-sample QPCR verification of differential expression of the LOC101929707 gene. According to the sample collection method in Example 1, 100 blood samples from normal subjects and 100 patients with ischemic stroke were selected.

[0063] 2. The RNA extraction steps are the same as in Example 1.

[0064] 3. Reverse transcription:

[0065] (1) Reverse transcription reaction

[0066] Use 25μl reaction system, take 1μg total RNA for each sample as template RNA, and add the following components to PCR tubes: DEPC water, 5× reverse transcription buffer, 10mM dNTP, 0.1mM DTT, 30μM Oligo dT, 200U / μl M-MLV, template RNA. Incubate at 42°C for 1 hour, then centrifuge briefly at 72°C for 10 minutes.

[0067] (2) Primer design

[0068] The primer sequence of LOC101929707 gene is:

[0069] Forward primer: 5'-AATGGTTCTGGATGGATTA-3' (SEQ ID NO.2)

[0070] Reverse primer: 5'-T...

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Abstract

The invention discloses application of lncRNA (long non-coding ribonucleic acid) to diagnosing cerebral ischemic stroke. The lncRNA is LOC101929707. The application has the advantages that correlation between LOC101929707 genes and occurrence and development of the cerebral ischemic stroke is discovered, and accordingly the LOC101929707 can be used as a possible detection target to be applied to early diagnosing the cerebral ischemic stroke on the basis of the correlation.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the application of lncRNA in the diagnosis of ischemic stroke, specifically the lncRNA is LOC101929707. Background technique [0002] Stroke, also known as apoplexy or cerebrovascular accident, is a group of acute cerebrovascular diseases with a common feature of focal neurological deficits caused by sudden onset of local blood circulation disorders in the brain. Due to its high morbidity, high mortality, high disability rate, and high recurrence rate, it has become one of the three major death diseases in the world. In my country, it has jumped to the first cause of death of urban residents, the second cause of death of rural residents, and the first among adults. disabling disease. In the past 20 years, with the increase of the natural population and the impact of aging, the incidence and mortality of stroke in my country have been on the rise. According to statistics, there are more th...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/113
CPCC12Q1/6883C12Q2600/178
Inventor 何文贞江耀睦陈荣波
Owner THE FIRST AFFILIATED HOSPITAL OF SHANTOU UNIV MEDICAL COLLEGE
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