SNP loci associated with aortic dissection disease and its application

A technology of aortic dissection and site, applied in the field of biomedical detection, can solve problems such as difficult early risk assessment, high AD vascular risk, and limited positive rate

Active Publication Date: 2022-07-15
王赞鑫
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] AD has a high vascular risk and is not easy to detect early. The positive rate of genetic diagnosis based on known disease-causing genes is limited, making early risk assessment difficult, and it is urgent to discover new molecular markers

Method used

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  • SNP loci associated with aortic dissection disease and its application
  • SNP loci associated with aortic dissection disease and its application
  • SNP loci associated with aortic dissection disease and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Sample Collection

[0030] From January 2017 to April 2018, 99 patients with Stanford type A aortic dissection were diagnosed by aortic CTA in Sun Yixian Cardiovascular Hospital of Shenzhen, and 2 mL of whole blood and 590 normal control samples were collected from the database searched by Mykino Sequencing Company . The informed consent of the patients has been obtained and reviewed by the ethics committee.

[0031] Sample processing: Mix EDTA anticoagulated whole blood with Trizol at a ratio of 1:1, mix well and place in a 1.8mL cell cryopreservation tube, quickly cool in liquid nitrogen for 30s, and store in a -80°C refrigerator.

Embodiment 2

[0032] Example 2 Extraction of DNA from blood samples

[0033] (1) Add 1 mL of CL cell lysate to 1 mL of blood anticoagulated by EDTA (0.01M, China, Huamei Bioengineer), gently invert and mix 6 times, centrifuge at 3600 rpm for 5 min, and discard the supernatant;

[0034] (2) Pour 1 mL of CL cell lysate into the centrifuge tube again, gently invert and mix 6 times, centrifuge at 3600 rpm for 5 minutes, and discard the supernatant; on the premise of ensuring that the precipitate remains in the tube, invert the centrifuge tube in Let stand on clean absorbent paper for 2min;

[0035] (3) configure the mixture of proteinase K and buffer FG;

[0036] (4) Add 500 μL of the mixture of proteinase K and buffer FG, and then mix until the solution has no clumps;

[0037] (5) 65℃ water bath for 30min, invert and mix several times during this period;

[0038] (6) Add 1 mL of isopropanol, then invert and mix until clustered or filamentous genomic DNA appears;

[0039] (7) Centrifuge at ...

Embodiment 3

[0047] Example 3 Whole exome sequencing

[0048] 1. DNA library construction

[0049] 3 μg of DNA was fragmented by ultrasonic, the ends of the fragments were blunted, A was added at the 3' end, and the adaptor was connected, and fragments between 350 and 400 bp were selected to prepare a whole genome library. The library samples were quality controlled using an Agilent 2100 Bioanalyzer (Agilent Technologies, USA).

[0050] 2. Target region capture and sequencing

[0051] Whole exome detection was performed using GenCap liquid phase capture target gene technology (Beijing Mykino Co., Ltd.). Mix 1 μg of DNA library with BL buffer and probe, heat at 95 °C for 7 min, and heat at 65 °C for 2 min, add 23 μL of HY buffer preheated to 65 °C, and hybridize at 65 °C for 22 h. 50 μL of MyOne magnetic beads (Life Technology, USA) were washed 3 times with 500 μL of 1X binding buffer, and resuspended in 80 μL of 1X binding buffer. Add 64 μL of 2X Binding Buffer to the hybridization mix...

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Abstract

The present invention discloses a method for screening SNP sites related to aortic dissection disease based on whole exome sequencing technology. The SNP sites include one or two of the following aortic dissection susceptibility SNP sites: PTK6_c. 205G>A_p.E69K; TSHZ2_c.724C>T_p.R242C. Further, the present invention provides the application of the SNP site in a detection product for detecting aortic dissection disease. The present invention discovers two aortic dissection susceptible SNP loci and studies their application prospects in early prediction of aortic dissection, and the aortic dissection-related gene loci provided by the present invention may be early biomarkers of aortic dissection It provides a new direction for further studying the genetic molecular mechanism of aortic dissection and exploring drug targets for early prevention and treatment of aortic dissection.

Description

technical field [0001] The invention relates to the technical field of biomedical detection, in particular to a SNP site related to aortic dissection disease and its application. Background technique [0002] Aortic dissection (AD) refers to the rupture of the aortic intima under the action of a series of external factors (such as hypertension, trauma, etc.), based on the presence or absence of lesions in the aortic wall itself. The blood aortic intimal rupture invades the middle layer of the aortic wall, and the middle layer of the aorta is continuously torn and separated longitudinally, resulting in a state of coexistence of true and false aortic lumen. The onset of aortic dissection is rapid, the development is rapid, and the prognosis is dangerous. Despite the continuous improvement of various current treatments, the morbidity and mortality of AD remains high, and if not properly and timely treated, AD mortality is extremely high. It has been reported that nearly 20% o...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883C12Q1/6869G16B30/10
CPCC12Q1/6883C12Q2600/156C12Q1/6869C12Q1/6837G16B20/20G16B30/00G16B40/00C12Q2600/106C12Q2531/113C12Q2565/131C12Q2535/122C12Q2523/301
Inventor 王赞鑫魏民新
Owner 王赞鑫
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