Cerebrospinal fluid VGF protein kit and application thereof in medulloblastoma metastasis evaluation

A technology for medulloblastoma and cerebrospinal fluid, applied in the biological field, can solve problems such as the inability to detect differential proteins, decrease the stability and sensitivity of quantitative analysis, and achieve high clinical application value

Pending Publication Date: 2020-11-17
AFFILIATED HUSN HOSPITAL OF FUDAN UNIV
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  • Application Information

AI Technical Summary

Problems solved by technology

Facts have proved that the ionization degree of different peptides and the response of different ions in the mass spectrometer are not the same, so the use of information of each peptide in traditional mass spectrometry to deduce the content of the corresponding protein may lead to a decrease in the stability and sensitivity of quantitative analysis. The consequence is that some differential proteins cannot be found

Method used

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  • Cerebrospinal fluid VGF protein kit and application thereof in medulloblastoma metastasis evaluation
  • Cerebrospinal fluid VGF protein kit and application thereof in medulloblastoma metastasis evaluation

Examples

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Embodiment 1

[0023] Analysis of peptide content in cerebrospinal fluid by DIA method

[0024] CSF sample preparation: Cells were centrifuged at 20,000g for 10 minutes at 4°C in a refrigerated centrifuge to remove insoluble material and cells. Add 1M DTT to each case of cerebrospinal fluid to a final concentration of 5mM, and reduce at 56°C for 1 hour. After cooling to room temperature, 0.5M IAM was added to a final concentration of 10 mM, and alkylation was carried out at room temperature for 45 minutes. The final concentration of L-cysteine ​​at room temperature is 20mM, and the alkylation reaction is terminated for 20 minutes; the final concentration of 1M TEAB is added to 0.1M, and trypsin (trypsin: sample, 1:50, m / m) is enzymatically hydrolyzed overnight , add trypsin again for 4 hours the next day; add formic acid to a final concentration of 1% to stop the enzymatic hydrolysis; extract the product with a C18 extraction column, dry and extract it with a vacuum concentrator after desal...

Embodiment 2、DI

[0029] Embodiment 2, DIA method detects cerebrospinal fluid VGF protein concentration

[0030] Collection of specimens: After obtaining the consent of 29 patients with medulloblastoma (including 14 patients with no metastasis and 15 patients with distant metastasis) who visited the Department of Neurosurgery of Huashan Hospital from 2006 to 2014, they underwent routine lumbar puncture. 5ml of cerebrospinal fluid was collected from each person and stored in a -80°C refrigerator;

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Abstract

The invention belongs to the technical field of biology, and relates to a tumor diagnosis kit, in particular to a cerebrospinal fluid VGF protein kit and application of the cerebrospinal fluid VGF protein kit to detection and pre-judgment of myeloblastoma metastasis. The kit comprising mouse anti-human VGF biotin labeled monoclonal antibody, streptomycin antibiotic protein peroxidase working fluidand other components is designed, then a collected cerebrospinal fluid sample is detected through an ELISA method, corresponding diagnosis is conducted according to the concentration of VGF in cerebrospinal fluid, and when the concentration of VGF is reduced to one third of the concentration of VGF after an operation, it is preliminarily judged that myeloblastoma cerebrospinal fluid metastasizes.The method disclosed by the invention is applied to diagnosis of myeloblastoma prediction, can be used for screening high-risk groups and guiding tumor staging, and provides a basis for searching sensitive and specific myeloblastoma cerebrospinal fluid markers.

Description

technical field [0001] The invention belongs to the field of biological technology, and relates to a tumor diagnosis kit, in particular to a cerebrospinal fluid VGF protein kit and its application in evaluating medulloblastoma metastasis. The kit detects the VGF protein in the cerebrospinal fluid, and provides a basis for searching for sensitive and specific cerebrospinal fluid markers of medulloblastoma metastasis. Background technique [0002] The prior art discloses that medulloblastoma is the most common malignant brain tumor in children, accounting for about 20% of children's tumors. The prognosis of this disease in adults is also very poor. Medulloblastoma has become a major brain tumor that endangers people's health. disease. In 2016, WHO classified medulloblastoma into different molecular subtypes: SHH, WNT, GROUP3, and GROUP4. The current standard treatment for medulloblastoma is: surgical resection + postoperative radiotherapy + postoperative chemotherapy. The 5-...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/6818G01N33/6848
Inventor 姚瑜王玉元钟平唐超李捷
Owner AFFILIATED HUSN HOSPITAL OF FUDAN UNIV
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