Serum marker for diagnosing intracranial aneurysm and serum marker for predicting rupture potential of intracranial aneurysm

A technique for intracranial aneurysm and serum markers, which is applied in the field of medicine and biology to achieve the effects of reducing inspection costs and high sensitivity

Pending Publication Date: 2021-06-11
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Currently, there are no clinically available biomarkers fo

Method used

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  • Serum marker for diagnosing intracranial aneurysm and serum marker for predicting rupture potential of intracranial aneurysm
  • Serum marker for diagnosing intracranial aneurysm and serum marker for predicting rupture potential of intracranial aneurysm
  • Serum marker for diagnosing intracranial aneurysm and serum marker for predicting rupture potential of intracranial aneurysm

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0112] Collection and preparation of 212 cases of ruptured intracranial aneurysm / unruptured intracranial aneurysm serum and normal human serum

[0113] Randomly grouped according to gender and age, 100 healthy people (NC) were collected as controls, 57 patients with unruptured intracranial aneurysm (UR), and 55 patients with ruptured intracranial aneurysm (R).

[0114] Main instruments and reagents: blood collection tubes, centrifuge tubes, BCA quantitative kit (Thermo Fisher Scientific, USA), peptide quantitative kit (Thermo Fisher Scientific, USA), tetraethylammonium bromide, DTT, IAA, trypsin, etc. .

[0115] Experimental steps:

[0116] 1. Collect serum samples:

[0117] (1) Collect whole blood into vacuum blood collection tubes, gently mix up and down for 5-6 times;

[0118] (2) Place at 4°C for 30-45 minutes;

[0119] (3) Centrifuge at 1000g for 10min at 4°C (after centrifugation, the upper layer of serum, the middle gel, and the lower layer of blood cells);

[0120...

Embodiment 2

[0129] Targeted proteomic validation of serum markers of ruptured / unruptured intracranial aneurysms

[0130] The expression levels of 20 candidate protein factors (25 peptide factors) in 212 serum samples were determined by PRM mass spectrometry.

[0131]Main instrument: Orbitrap Exploris 480Mass Spectrometer (Thermo Fisher Scientific, USA)

[0132] Experimental procedures: Chromatographic column model: 50cm C18 chromatographic separation column (2μm, 75μm, x 500mm, Thermo Fisher Scientific, USA); flow rate: 200nL; gradient: phase A: 0.1% FA aqueous solution; phase B: 80% ACN. Separation gradient 0-50% phase B, total separation time: 65min; Orbitrap Exploris 480 mass spectrometry parameters: use PRM mode for collection. Primary scan range (m / z): 350-1600; import the mass-to-charge ratio and retention time information (Table 1) of the target peptide into the designated list.

Embodiment 3

[0134] PRM data collection and bioinformatics analysis

[0135] Import the original data into Skyline-Daily software for data collection: (1) import the Fasta file of the target protein into skyline; (2) peptide setting: the background library is the Uniprot library, and the search result file is imported as the spectral library, and the peptide length is 8-25AA; (3) Product ion (Transition) extraction settings: parent ion charge: 2; select the strongest product ion for extraction according to the spectral library; the acquisition mode is Targeted; 25 peptides selected by 20 candidate protein factors The parent and child ion information of the segment is shown in Table 2. (4) Manually check the extraction time of each product ion and the first-level ID time, and compare the retention time, peak area, etc.; (5) Export the peptide ratio results for subsequent statistical analysis.

[0136] The exported data were statistically analyzed with SPSS Statistics 20 software: Mann-Whit...

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Abstract

The invention relates to a serum marker for diagnosing intracranial aneurysm and a serum marker for predicting rupture potential of intracranial aneurysm. The serum marker for diagnosing intracranial aneurysm is selected from one or more of the following protein factors: PRTN3, CTSG, PDLIM1, MMP9, IGKV3-20, MPO, PROC, LTF, FGA, FGB, PPBP or SERPINA1. The serum marker for predicting the rupture potential of the intracranial aneurysm is selected from one or more of the following protein factors: SAA1, LRG1, FGL1, PTGDS, COMP, IGKV4-1, APOA4, FGA, FGB and FGG. Compared with the prior art, the serum marker for diagnosing the intracranial aneurysm and the serum marker for predicting rupture of the intracranial aneurysm are screened by applying a PRM mass spectrum technology based on an Orbitrap Explores 480 mass spectrometer and a bioinformatics method, so that a new thought is provided for clinical diagnosis and treatment of the intracranial aneurysm; and high sensitivity, specificity and accuracy are achieved. In addition, the protein mass spectrum technology can be perfected and improved, and the inspection cost is reduced, so that the serum marker is enabled to be more suitable for clinical application of precision medicine.

Description

technical field [0001] The invention belongs to the technical field of medicine and biology, and in particular relates to a serum marker for diagnosing an intracranial aneurysm and a serum marker for predicting the rupture potential of an intracranial aneurysm. Background technique [0002] Intracranial aneurysm (Intracranial aneurysm, IA) is a complex multifactorial cerebrovascular disease. Intracranial aneurysms occur in 2-3% of the general population, and about 1% of intracranial aneurysms rupture every year. Intracranial aneurysm rupture is the most common cause of aneurysmal subarachnoid hemorrhage (aSAH), with a mortality rate of 35% for the first hemorrhage and 60% to 80% for the second hemorrhage. Most survivors have obvious neurological impairment. The pathogenesis of intracranial aneurysms is not clear, and it may be related to factors such as hypertension, smoking, alcohol consumption, lipid accumulation, and hemodynamics. [0003] Currently, the diagnosis and s...

Claims

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Application Information

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IPC IPC(8): G01N33/574
CPCG01N33/57407G01N33/57488
Inventor 刘晓慧熊悦婷郑永涛冷冰杨芃原
Owner FUDAN UNIV
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