Application of human circular RNA in preparation of heart disease-related products
A technology of heart disease and related products, applied in the field of biomedicine, can solve the problems of unfavorable diagnosis, specificity, poor sensitivity and stability, etc., and achieve the effect of good detection sensitivity and specificity, significant changes, and high stability
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Embodiment 1
[0055] Identification of circHED in human blood:
[0056] CircHED in blood was identified by inverse PCR. Take 500 μl of human blood, add 500 μl Trizol (Takara RNAiso Plus 9108), turn the centrifuge tube upside down several times, let it stand at room temperature for 15 minutes, extract total RNA according to the instructions, and further use the Takara RR0037A reverse transcription kit to pair random primers Random 6-mer RNA was reverse-transcribed to synthesize cDNA; another 300 μl of blood was extracted through the TIANGEN Blood Genomic DNA Extraction System (DP349) to extract genomic DNA (gDNA). Using blood cDNA or gDNA as a template, the sequence of the circHED terminal junction site was amplified by PCR using the reverse primer, and the forward primer was used as a control. The amplified products were detected by agarose gel electrophoresis, and the amplified products using cDNA as a template were subjected to Sanger sequencing.
[0057] The primer sequences are as fol...
Embodiment 2
[0067] Detection of circHED in the blood of patients with myocardial infarction:
[0068] In this experiment, 50 blood samples were collected from normal people with no history of cardiovascular disease and 50 patients with acute myocardial infarction. Patients with myocardial infarction should meet the following clinical manifestations: (1) angina pectoris lasted for more than 30 minutes; (2) ECG showed typical myocardial infarction features; 3) Increased levels of molecular markers such as creatine kinase, lactate dehydrogenase, and troponin T or I in the blood. Blood RNA was extracted and cDNA was synthesized according to the method described in Example 1, and 18SrRNA was used as an internal reference, and qPCR was used for detection, and 2 -ΔΔCT The relative expression level of circHED in blood was calculated by the method. Novazan ChamQ Universal SYBR qPCR qPCR Master Mix was used in the qPCR kit, and the experimental operation was performed according to the instructions...
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