Circular RNA marker for detecting cerebral ischemia-reperfusion injury and application
A technology of cerebral ischemia-reperfusion and molecular markers, applied in the direction of DNA/RNA fragments, recombinant DNA technology, biochemical equipment and methods, etc., can solve the problems of complex pathogenesis and multiple influencing factors of cerebral ischemia-reperfusion injury , to achieve the effect of improving clinical treatment effect, quantitative accuracy, good application value and prospect
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Embodiment 1
[0023] Example 1 Total RNA extraction, circRNA sequencing and screening
[0024] Prepare rat suture embolization MCAO / R cerebral ischemia-reperfusion injury model, collect peripheral blood, select 3 groups of peripheral blood samples for inspection, and use peripheral blood samples of normal rats as a control for RNA extraction and sequencing;
[0025] The specific method is as follows: 1. According to the instructions, use TRIZOL reagent from Thermo Fisher to extract 5 μg Total RNA from each sample, then use Epicentre Ribozero rRNA Removal Kit to remove rRNA kit, and use RNase R to digest the linear RNA in it , using Illumina’s directional library preparation kit to generate a sequence library. After cluster generation, it was sequenced on a high-throughput 4000 sequencing platform to generate 150-bp double-ended reads. The above sequencing and circRNA / mRNA analysis were all performed by Novogene source company.
[0026] 2. RT-qPCR verification
[0027] Use the RevertAid Fi...
specific Embodiment 2
[0035] Specific Example 2 Identification and detection of three corresponding circular circRNA 0002086, circRNA0001606 and circRNA 0002023 in human cerebral ischemia-reperfusion blood.
[0036] Methods: In this example, 28 peripheral blood samples from patients with cerebral ischemia-reperfusion injury and 28 samples from normal subjects were collected, and the levels and specificities of circRNA 0002086, circRNA 0001606 and circRNA 0002023 were detected.
[0037] It was found that, if Figure 2-3 As shown, the expression level of circRNA 0002086 in patient samples was significantly higher than that in the normal group, the difference was statistically significant (P<0.001), and the specificity of circRNA 0002086 as cerebral ischemia-reperfusion injury was 87%.
[0038]The results showed that in the blood of patients with cerebral ischemia-reperfusion injury, the expression level of circRNA 0002086 was also significantly higher than that of healthy people. Therefore, circRNA 0...
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