A lncRNA marker and its application in the diagnosis of acute myocardial infarction
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A kind of acute myocardial infarction, primer pair technology, applied in the field of biomedicine
Active Publication Date: 2020-10-30
青岛山大齐鲁医院(山东大学齐鲁医院(青岛))
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At present, there are few studies on long-chain non-coding RNA in myocardial infarction, and compared with traditional biochemical markers, long-chain non-coding RNA has a high degree of stability. Long non-coding RNA is of great significance for early diagnosis of myocardial infarction
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Embodiment 1
[0027] 1. Processing of plasma samples
[0028] The collected blood was refrigerated at 4°C for 30 minutes, then centrifuged at 3000 rpm for 15 minutes at room temperature, and the upper layer of plasma was transferred to a 1.5ml centrifuge tube and stored in a -20°C refrigerator for later use.
[0029] 2. Extraction of plasma total RNA
[0030] (1) Thaw the separated plasma on ice, take 250μl plasma sample into a 2ml centrifuge tube, add 750μl Trizol, mix well by pipetting, and let stand for 5min;
[0031] (2) Add 200 μL of chloroform, mix upside down 20 times, let stand at room temperature for 5 minutes, centrifuge at 12000 rpm for 15 minutes;
[0032] (3) Carefully transfer the upper aqueous phase to a new RNase-free centrifuge tube, add an equal volume of isopropanol, mix well and place on ice for 10 minutes;
[0033] (4) Centrifuge at 13,000 rpm, 4°C for 10 minutes, discard the supernatant, and keep the precipitate;
[0034] (5) Add 500 μL pre-cooled 70% ethanol to dis...
Embodiment 2
[0062] A fluorescent quantitative PCR kit for diagnosing acute myocardial infarction
[0063] 1. Composition
[0064] LINC01104 primer pair, GAPDH primer pair, SYBR Green fluorescent dye, and ddH2O.
[0065] LINC01104
[0066] Forward primer 5'-AGGGCATGACAGGCAATTCA-3'
[0067] Reverse primer 5'-AGTGATCCTCTTGCCTTGGC-3'
[0068] GAPDH
[0069] Forward primer 5'-ACGTGTCAGTGGTGGACCTG-3'
[0070] Reverse primer 5'-GTGTAGCCCAGGATGCCCTT-3'
[0071] 2. How to add
[0072]
[0073] 2. Reaction conditions
[0074] Reaction conditions: 95°C for 10min; 95°C for 15s, 60°C for 60s for 40 cycles.
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Abstract
The invention provides an lncRNA marker and application thereof in acute myocardial infarction diagnosis. The LncRNA marker is LINC01104, and the expression quantity of the LINC01104 in an acute myocardial infarction patient is lower than that of a normal person. The invention further discloses a fluorescent quantitative PCR kit for detecting acute myocardial infarction. The kit contains a reagentfor measuring expression of the LINC01104. Compared with a traditional biomarker, the LINC01104 marker has better tissue specificity and good stability, and the sensitivity and specificity of acute myocardial infarction diagnosis can be greatly improved.
Description
technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to the application of a LncRNA marker in the diagnosis of acute myocardial infarction. Background technique [0002] Acute myocardial infarction is an acute cardiovascular disease with high mortality, high disability rate and high medical expenses. At present, acute myocardial infarction has become a prominent public health and social problem. Acute myocardial infarction is based on coronary atherosclerosis, and the rupture of plaques often leads to thrombosis in the official cavity and vascular obstruction, followed by myocardial necrosis due to interruption of coronary blood flow. In my country, with the change of lifestyle and diet structure, the incidence of AMI is showing an increasing trend year by year. At present, the biomarker widely used in clinical practice for the diagnosis of myocardial infarction is troponin (cTnI), but the concentration of troponin in...
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