Biomarker for detecting occlusion or stenosis of coronary artery and preparation method thereof, and reagent kit containing biomarker

A biomarker, coronary artery technology, applied in biochemical equipment and methods, microbial determination/examination, DNA/RNA fragments, etc., can solve problems such as affecting accuracy, and achieve ideal accuracy, high sensitivity and specificity. Good results

Active Publication Date: 2017-05-10
HEBEI MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, factors that are difficult to overcome, such as motion artifacts and coronary artery calcification, often affect the accuracy of this evaluation method

Method used

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  • Biomarker for detecting occlusion or stenosis of coronary artery and preparation method thereof, and reagent kit containing biomarker
  • Biomarker for detecting occlusion or stenosis of coronary artery and preparation method thereof, and reagent kit containing biomarker
  • Biomarker for detecting occlusion or stenosis of coronary artery and preparation method thereof, and reagent kit containing biomarker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1c

[0056] Preparation of embodiment 1circ-Sirt1 gene

[0057] 1) Extraction of total RNA in human plasma: extraction using Trizol LS Reagent reagent based on Life Technologies.

[0058] (a) Using EDTA-K 2 Anticoagulant tubes were used to collect peripheral venous blood samples. Within 2 hours of the collected blood sample, centrifuge at 2000g at 4°C for 20min, collect the supernatant and centrifuge again at 10000g at 4°C for 20min, and store the supernatant at -80°C.

[0059] (b) Add 900 μl Trizol LS to 300 μl plasma, mix well and lyse at room temperature for 5 minutes.

[0060] (c) Add 0.2ml of chloroform:isoamyl alcohol (24:2), shake for 15s, and let stand at room temperature for 2min.

[0061] (d) Centrifuge at 12000 g for 15 min at 4°C, and take the supernatant.

[0062] (e) Add 0.6ml of isopropanol, mix the liquid in the tube gently, and let stand at -20°C for 2h.

[0063] (f) Centrifuge at 12000 g for 15 min at 4°C, and take the supernatant.

[0064] (g) Add 1 ml of 7...

Embodiment 2

[0076] Example 2 Using the biomarkers of the present invention to detect and evaluate the degree of stenosis of the coronary arteries of patients

[0077] Blood samples: Plasma samples from 20 patients with coronary atherosclerosis came from inpatients in the Department of Cardiology, The Second Affiliated Hospital of Hebei Medical University. Plasma samples from 20 healthy individuals were obtained from the Physical Examination Center of the Second Affiliated Hospital of Hebei Medical University.

[0078] 1) Inclusion criteria

[0079] Patients with initial or reoccurrence of coronary artery sclerosis occlusive stenosis obtained by coronary angiography.

[0080] 2) Exclusion criteria

[0081] Healthy people: healthy people of similar age, no history of coronary heart disease, no history of surgery, normal blood lipids, blood sugar, blood pressure, no family genetic diseases, no liver and kidney insufficiency;

[0082] 3) Grouping of collected specimens

[0083] (a) Experi...

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Abstract

The invention discloses a biomarker for detecting occlusion or stenosis of coronary artery and a preparation method thereof, and a reagent kit containing the biomarker. The name of the biomarker is circ-Sirtl, and the biomarker comprises a nucleotide sequence gene shown in SEQ ID NO: 1. The preparation method comprises the following steps of (A) extracting total RNA (ribonucleic acid) in human blood plasma; (B) removing remained DNA (deoxyribonucleic acid) in the total RNA; (C) reversely transcribing into cDNA by a random primer; (D) designing a primer, and performing PCR (polymerase chain reaction) amplifying, cloning and sequencing. The reagent kit comprises a blood plasma total RNA extraction reagent set, a RNA reverse transcription reagent set, a real-time fluorescent quantitative PCR reagent set, a probe set, and a real-time fluorescent quantitative PCR primer set. The circ-Sirtl biomarker and the detection regent kit have the outstanding effects of high sensitivity and strong specificity in detecting and evaluating the stenosis degree of the coronary artery of a patient.

Description

technical field [0001] The invention relates to a biomarker for diagnosing diseases, a diagnostic kit and a preparation method of the marker. Specifically, it is a biomarker that can be used to detect coronary artery occlusion stenosis, a kit containing the biomarker and a preparation method of the biomarker. Background technique [0002] Coronary artery stenosis belongs to coronary artery disease and is an important cause of coronary heart disease. Accurate assessment of the degree of coronary artery stenosis is extremely important for early detection and intervention of coronary heart disease. At present, coronary angiography (CAG) is the gold standard for clinical examination of coronary artery lesions, but its examination has the disadvantages of trauma, high cost, and long time. In recent years, with the development of CT technology, multi-slice CT coronary angiography has been more and more used clinically. Multi-slice CT coronary angiography can display plaque comp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6883C12Q2600/158
Inventor 韩梅孔鹏于媛王海月张旭慧
Owner HEBEI MEDICAL UNIVERSITY
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