Method of rapidly detecting accumulation of tumor tissue 2-HG

Abstract

The invention belongs to the technical field of biology, and relates to a method of rapidly detecting the accumulation of the carcinogenic metabolite of tumor cells namely 2-hydroxyl glutaric acid (2-HG). The method comprises the following steps: adding an obtained tumor cell sample in a centrifuge tube, adding pre-cooled pure methanol, grinding the sample by a homogenizer, allowing the solution to stand still, subjecting the solution to centrifugation (14000 rpm), transferring the supernate to a glass bottle, fully drying the supernate in a vacuum drying centrifuge, adding a derivatization reagent (MTBSTFA), placing the sample at a temperature of 70 DEG C, cooling to the room temperature, and loading the sample into Agilent 7890A-5975C, wherein the chromatographic column is HP-5MS, the carrier gas is highly pure helium, and a NIST database search technology and a standard substance verification method are combined to identify 2-HG. The provided method can be used to judge whether highlevel accumulation of 2-HG exists in brain tissues or not during a surgery process; so IDH1 mutant glioma can be rapidly identified under the assistance of the provided method, and important information is provided for the individual treatment after surgery.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to carcinogenic metabolite 2-hydroxyglutarate (2-HG), in particular to a method for rapidly detecting the accumulation of 2-HG in tumor tissue. By detecting the accumulation of 2-HG in tumor tissue, it helps For the rapid diagnosis of IDH1 mutations. Background technique [0002] The prior art discloses that glioma is the most common tumor of the central nervous system, among which the cure rate of malignant glioma is low, the recurrence rate is high, and the prognosis is extremely poor. At present, synchronous radiotherapy and chemotherapy after surgery plus 6 courses of treatment are mostly used clinically Temozolomide (TMZ) chemotherapy, the average survival period is only 12-15 months. Therefore, the current clinical problem to be solved for malignant glioma is to find a method for early diagnosis and early treatment. [0003] In current clinical practice, stereotaxic biopsy is requ...

AI Technical Summary

Problems solved by technology

[0003] In current clinical practice, stereotaxic biopsy is required for some lesions that are not clearly diagnosed. Typical cases are easier to diagnose. However, in practice, the pathology of frozen sections during surgery is often not typical, and pathologists can only observe gliosis. And gliosis can appear in low-grade glioma, normal brain tissue, inflammation, some lymphoma, etc.

Although mutant IDH1 immunohistochemical staining can distinguish glioma from other conditions, it usually takes 5-7 days, which takes a long time, and intraoperative judgment cannot be realized, and sometimes even repeated punctures are required, which increases the burden on patients and the risk of surgery