Application of VMP1 gene in pathological diagnosis of glioblastoma and preparation of kit thereof

A glioblastoma and diagnostic kit technology, applied in the field of medicine, can solve the problems of low expression level and low expression of VMP1 gene, achieve timely diagnosis, high accuracy rate, and broad application prospects

Pending Publication Date: 2019-10-22
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Existing studies have shown that the expression level of VMP1 gene in normal human brain tissue is extremely low, and the expression level is also low in WHO grade Ⅱ and Ⅲ gliomas.

Method used

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  • Application of VMP1 gene in pathological diagnosis of glioblastoma and preparation of kit thereof
  • Application of VMP1 gene in pathological diagnosis of glioblastoma and preparation of kit thereof
  • Application of VMP1 gene in pathological diagnosis of glioblastoma and preparation of kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Application of VMP1 / TMEM49 gene mRNA as a molecular pathological diagnostic marker in the diagnosis of glioblastoma.

[0062] 1. Collection and processing of glioma tissue samples and extraction of total RNA.

[0063] (1) Tumor pathological tissues of patients with glioma were collected during the operation.

[0064] (2) After flushing with normal saline, the pathological tissues were frozen in liquid nitrogen.

[0065] (3) Cut 100 mg of each sample into an EP tube and add 1 ml of Trizol reagent for homogenization.

[0066] (4) Add 500 μL of chloroform, shake well and centrifuge at 12000 rpm for 5 min at 4°C.

[0067] (5) Carefully suck out the uppermost layer of liquid into a new EP tube, add 500 μL of isopropanol, mix well, let stand for 10 minutes, and centrifuge at 12,000 rpm for 5 minutes at 4°C.

[0068] (6) Discard the supernatant, add 1 mL of 75% ethanol (prepared with DEPC water), and centrifuge at 12000 rpm for 1 min at 4°C. Repeat this step one more time....

Embodiment 2

[0083] Application of VMP1 protein as a molecular pathological diagnostic marker in the diagnosis of glioblastoma.

[0084] 1. Preparation of tissue slices

[0085] (1) The pathological tissue of glioma cryopreserved in liquid nitrogen was taken out for frozen embedding and sectioning.

[0086] (2) Flatten the frozen section on the pathological glass slide.

[0087] 2. Immunohistochemical staining

[0088] (1) Add 200 μl immunohistochemical non-specific antigen blocking solution (containing 1% calf serum and 0.3% Triton-100) to the tissue section on the slide, and incubate on a horizontal shaker at room temperature for 2 hours.

[0089] (2) Use primary antibody diluent to prepare VMP1 antibody solution (1:200). After aspirating the blocking solution, add 200 μl of antibody solution dropwise to the tissue section, so that the section is completely submerged in the liquid. Place the slides in a wet box on a horizontal shaker and incubate overnight at 4°C.

[0090] (3) After...

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Abstract

The invention discloses application of a VMP1 gene as a glioblastoma pathological diagnosis marker, and also discloses a glioblastoma diagnostic kit. The high expression of a VMP1 gene product in theglioblastoma has very high specificity, so that the WHO grade IV glioblastoma is effectively and definitely diagnosed; a basis can be provided for the definite diagnosis and treatment of the clinicalglioblastoma, and it is expected to improve the prognosis of a patient by performing related treatment as soon as possible; by combining a PCR method with a protein immunohistochemical staining method, compared with a traditional method, the VMP1 gene product has the advantages of low cost, high speed, high sensitivity, high accuracy and the like, and can meet needs of large-scale experiments andclinical application; the molecular marker performs diagnosis more timely and more specifically, and has broad application prospects.

Description

Technical field: [0001] The invention relates to the technical field of medicine, in particular to the application of VMP1 gene in pathological diagnosis of glioblastoma and the preparation of a kit thereof. Background technique: [0002] Glioblastoma is the most malignant glioma, which belongs to WHO grade IV. In the United States, the incidence rate of malignant glioma is 5.26 per 100,000 residents, and about 17,000 new cases are diagnosed every year; malignant glioma in China The incidence rate is also relatively high, about 3-6 people per 100,000 people are diagnosed with malignant glioma, and as many as 30,000 people lose their lives every year. The treatment of glioblastoma currently implements a multimodal treatment of surgical resection combined with radiotherapy and chemotherapy; however, even with such a comprehensive treatment approach, the prognosis of patients is still poor, the recurrence rate is extremely high, and the one-year survival rate is only 26% . A ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886G01N33/68G01N33/574
CPCC12Q1/6886G01N33/68G01N33/57407C12Q2600/118G01N2333/46
Inventor 祝刚杨倩
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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