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Pulmonary thromboembolism gene panel kit and application thereof

A technology for embolism and pulmonary thrombosis is applied in the field of liquid-phase gene panel capture kits for detecting pulmonary thromboembolism genes, which can solve the problems of inability to detect genes at the same time, slow detection speed and low efficiency, and achieve the effect of improving the depth of sequencing

Pending Publication Date: 2019-04-19
SHENZHEN PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The current routine detection of pulmonary embolism risk or pathogenic factors is mostly through Elisa (four blood coagulation items, etc.), gene detection chip or PCR and other genotyping technologies. The detection speed is slow, the efficiency is low, and the cost is high. The throughput of detection Low, it is impossible to detect a large number of genes at the same time, and to detect multiple indicators at the same time requires a variety of technical means

Method used

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  • Pulmonary thromboembolism gene panel kit and application thereof
  • Pulmonary thromboembolism gene panel kit and application thereof
  • Pulmonary thromboembolism gene panel kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Genomic DNA Sample Extraction

[0044] Use QIAamp DNA blood mini kit (QIAGEN, Cat: Y5-51106) to extract and purify DNA from venous blood of subjects, the specific steps are as follows:

[0045] 1. Take venous blood, centrifuge at 1000g for 15min at room temperature, absorb 1ml of blood cells, add 3ml of DNA lysate, mix thoroughly with a vortex shaker, and mark the name and date.

[0046] 2. Pipette 20 μl proteinase K to the bottom of a 1.5ml centrifuge tube.

[0047] Add 200 μl blood sample to the centrifuge tube, then add 200 μl Buffer AL to the sample, vortex for 15 seconds to mix.

[0048] 3. After incubating at 56°C for 10 minutes, centrifuge quickly to remove the droplets remaining in the cap of the 1.5ml centrifuge tube.

[0049] 4. Add 200 μl of absolute ethanol, and vortex for 15 seconds to mix. After shaking, quickly centrifuge, transfer the mixture to a QIAamp Mini centrifugal column, and centrifuge at 6000g for 1min.

[0050] 5. Place the QIAamp...

Embodiment 2

[0055] Example 2 Target gene whole exome capture, library construction

[0056] Design and synthesize the probe sequences, as shown in SEQ ID NO.1 to SEQ ID NO.184, and label with biotin, and prepare the gene Panel liquid chip.

[0057] The samples obtained in Example 1 were captured and detected by the all-exon gene panel liquid chip system of the present invention, and the target region library results captured before sequencing were obtained.

[0058] The gene panel liquid chip system of the present invention is used to efficiently enrich the DNA of the whole exon region of the target gene DNA in Example 1, and then perform high-throughput and high-depth sequencing on the Illumina Hiseq platform. The gene panel liquid phase chip system of the present invention is used in the library building and capture experiments. The selected supporting reagents and consumables are provided by the corresponding suppliers. The operations are all adjusted to the appropriate scheme accordin...

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Abstract

The invention relates to the technical field of biomedicine and particularly provides a liquid phase gene panel capture kit for detecting genes related to pulmonary thromboembolism and application thereof. The liquid phase gene panel capture kit comprises a capture probe covering 92 genes. The design of the gene panel capture kit integrates multi-level and multi-aspect genetic factors of pulmonaryembolism, is comprehensive in coverage and has obvious pertinence in target capture, genetic detection can be simultaneously conducted on genes with large fluxes, sequencing depth and coverage degreeof the genes can be effectively improved by efficiently enriching DNA of exon areas of target genes of people and adopting subsequent sequencing, and guarantee is provided for obtaining of more accurate gene genetic information and results.

Description

technical field [0001] The invention relates to the technical field of biomedicine, and specifically provides a liquid-phase gene panel (Panel) capture kit for detecting pulmonary thromboembolism genes and an application thereof. Background technique [0002] Pulmonary thromboembolism (PTE) refers to a disease caused by thrombus from the venous system or right heart blocking the pulmonary artery or its branches, with pulmonary circulation and respiratory dysfunction as the main clinical manifestations and pathophysiological features. PTE is a multifactorial disease caused by abnormal blood coagulation in the venous system, environmental and genetic factors. It is mainly manifested as thrombosis in the blood circulatory system. It has been reported that the incidence of DVT and PTE in the United States ranks third among cardiovascular diseases, second only to coronary heart disease and hypertension. At present, the epidemiology of PTE in my country lacks large-scale statist...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883
CPCC12Q1/6883C12Q2600/106
Inventor 傅应云乐雍建李博李亚珍韩雪梅黄启俊肖露李树林刘盛国赖胜敏
Owner SHENZHEN PEOPLES HOSPITAL
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