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Polypeptide segment diagnosis model and application thereof in prediction of myeloblastoma metastasis risk

A technology of medulloblastoma and peptides, which is applied in the diagnostic model of polypeptide segments and its application in predicting the risk of medulloblastoma metastasis, can solve the problem that differential proteins cannot be found, quantitative analysis stability and sensitivity decline, etc. question

Pending Publication Date: 2020-12-15
AFFILIATED HUSN HOSPITAL OF FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • 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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  • Polypeptide segment diagnosis model and application thereof in prediction of myeloblastoma metastasis risk
  • Polypeptide segment diagnosis model and application thereof in prediction of myeloblastoma metastasis risk
  • Polypeptide segment diagnosis model and application thereof in prediction of myeloblastoma metastasis risk

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Embodiment 1, research basis of the present invention

[0019] Analysis of peptide content in cerebrospinal fluid by DIA method

[0020] CSF sample preparation: Cells were centrifuged at 200009 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 was 20mM, and the alkylation reaction was terminated for 20 minutes; the final concentration of 1M TEAB was added to 0.1M, and trypsin (trypsin: sample, 1:50, m / m) was 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 ...

Embodiment 2

[0027] Embodiment 2, detection method of the present invention

[0028] 75 peptides that make up the diagnostic model

[0029] 1. CO5_HUMAN_LQGTLPVEAR_2_y7_1

[0030] 2. CO5_HUMAN_LQGTLPVEAR_2_y8_1

[0031] 3. ITIH4_HUMAN_TGLLLLSDPDK_2_precursor_2

[0032] 4. ITIH4_HUMAN_TGLLLLSDPDK_2_precursor[M+1]_2

[0033] 5. PRDX6_HUMAN_LPFPIIDDR_2_precursor[M+1]_2

[0034] 6. 7B2_HUMAN_SVNPYLQGQR_2_precursor_2

[0035] 7. 7B2_HUMAN_SVNPYLQGQR_2_precursor[M+1]_2

[0036] 8. 7B2_HUMAN_SVNPYLQGQR_2_precursor[M+2]_2

[0037] 9. 7B2_HUMAN_SVNPYLQGQR_2_y4_1

[0038] 10. 7B2_HUMAN_SVNPYLQGQR_2_y5_1

[0039] 11. 7B2_HUMAN_SVNPYLQGQR_2_y7_1

[0040] 12. 7B2_HUMAN_SVNPYLQGQR_2_y8_1

[0041] Thirteen, ANGT_HUMAN_VEGLTFQQNSLNWMK_2_y3_1

[0042] Fourteen, ANGT_HUMAN_VEGLTFQQNSLNWMK_2_y4_1

[0043] Fifteen, ANGT_HUMAN_VEGLTFQQNSLNWMK_2_y8_1

[0044] Sixteen, CA2D1_HUMAN_FFGEIDPSLMR_2_precursor[M+1]_2

[0045] Seventeen, CPVL_HUMAN_NNDFYVTGESYAGK_2_precursor_2

[0046] 18. CPVL_HUMAN_NND...

Embodiment 3

[0105] Example 3. Sensitivity and specificity of detecting cerebrospinal fluid B7-H4 concentration for diagnosing glioma by DIA method

[0106] Collection of specimens: Lumbar puncture was performed on 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. , 5ml of cerebrospinal fluid was collected from each person and stored in a -80°C refrigerator;

[0107] Detection of target concentration: thaw the cerebrospinal fluid sample in an ice bath for 30-60 minutes, centrifuge at 1500 rpm for 5 minutes, take the supernatant, and use a DIA machine according to the method in Example 2;

[0108] ROC curve analysis: using SPSS Statistics 20 software. Draw the ROC curve for diagnosing glioma (such as Figure 4 shown), the area under the curve is 0.9613, when the 37 peptides in the cerebrospinal fluid meet our predicted changes as the criteria ...

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Abstract

The invention belongs to the technical field of clinical tumor diagnosis, and particularly relates to a model and a prediction kit for predicting whether cerebrospinal fluid peptide fragments of a patient suffering from myeloblastoma are transferred or not in the future according to the change of the components of the cerebrospinal fluid peptide fragments. The method is applied to prediction of myeloblastoma metastasis, the sensitivity of the method is 70.8%, the specificity of the method is 100%, and the AUC value of the method is 0.9619. The method not only can guide tumor staging, but alsocan adjust the postoperative chemotherapy dosage according to the patient metastasis possibility and improve the patient prognosis. And a basis is provided for searching a sensitive and specific medulloblastoma cerebrospinal fluid marker.

Description

technical field [0001] The invention belongs to the field of tumor diagnosis and relates to a novel polypeptide segment medulloblastoma metastasis prediction kit. The kit detects 75 peptide segments in the cerebrospinal fluid and compares them with the cerebrospinal fluid after the initial operation. The method is applied to The prediction of medulloblastoma metastasis can not only guide tumor staging, but also adjust the dose of postoperative chemotherapy according to the possibility of patient metastasis, so as to improve the prognosis of patients. And provide a basis for finding sensitive and specific markers of medulloblastoma in cerebrospinal fluid. Background technique [0002] Medulloblastoma is the most common malignant brain tumor in children, accounting for about 20% of childhood tumors. The prognosis of adults is also very poor. Medulloblastoma is a serious brain disease that endangers people's health. In 2016, WHO classified medulloblastoma into different molecu...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/50
CPCG01N33/6848G01N33/5091G01N2560/00
Inventor 姚瑜王玉元钟平张振宇李捷唐超
Owner AFFILIATED HUSN HOSPITAL OF FUDAN UNIV
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