Glial fibrillary acidic protein (GFAP) antibody detection test kit, and application thereof

A detection technology for acidic proteins and antibodies, applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of no unified recommended method, and achieve the effect of convenient interpretation and high sensitivity

Pending Publication Date: 2020-02-18
THE SECOND AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the detection methods of glial fibrillary acidic protein antibody, that is, GFAP antibody, mainly in

Method used

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  • Glial fibrillary acidic protein (GFAP) antibody detection test kit, and application thereof

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Effect test

Embodiment 1

[0040] A glial fibrillary acidic protein antibody detection kit, which is based on rat brain tissue for detection, including rabbit-derived anti-human GFAP antibody, PBS buffer, fluorescent dye CY3, goat anti-rabbit IgG, FITC, goat anti-human IgG .

Embodiment 2

[0042] The application of the kit in the detection of glial fibrillary acidic protein antibody comprises the following steps:

[0043] Preparation of Frozen Rat Brain Tissue Sections

[0044] (1) Wistar adult rats were conventionally anesthetized, executed by decapitation, brain tissue was taken, and soaked in frozen 4% paraformaldehyde overnight;

[0045] (2) The cryostat was pre-cooled to -20°C, and the hippocampus and cerebellum of the rat brain tissue to be cut were embedded with OCT embedding agent;

[0046] (3) Slice according to the thickness of 30 μm, and soak the slice in PBS buffer solution;

[0047] (4) Carefully spread the cut slices on the glass slide;

[0048] (5) dry and set aside;

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Abstract

The invention provides a glial fibrillary acidic protein (GFAP) antibody detection test kit, and an application thereof. The application comprises the following steps: S1, preparing a frozen animal brain slice; S2, respectively sucking a cerebrospinal fluid to be detected and a diluted rabbit anti-human GFAP antibody, and dripping the cerebrospinal fluid to be tested and the diluted rabbit anti-human GFAP antibody on a reaction plate at a ratio of 1 to 1; S3, covering the reaction plate with the frozen animal brain slice which is turned over, and carrying out incubation overnight at a temperature of 4 DEG C; S4, rinsing the brain slice with a PBS buffer solution twice; S5, respectively sucking goat anti-rabbit IgG, which is diluted and marked by the water-soluble fluorescent dye CY3, and agoat anti-human IgG mixed solution, which is diluted and FITC-marked, and dripping the goat anti-rabbit IgG and the goat anti-human IgG mixed solution on the reaction plate at a ratio of 1 to 1; S6,covering the reaction plate with the frozen animal brain slice which is turned over, and carrying out two-hour incubation at the room temperature; S7, rinsing the brain slice with the PBS buffer solution twice; S8, sealing the slice; and S9, placing the sealed slice under a fluorescent microscope for observation. The test kit and the application thereof provided by the invention have the advantages that the sensitivity is higher than that of cell method based detection, and a result can be conveniently interpreted.

Description

technical field [0001] The invention relates to a glial fibrillary acidic protein antibody detection kit and application thereof. Background technique [0002] Glial fibrillary acidic protein astrocytopathy (GFAP) is the skeleton protein of astrocytes and also a marker protein. In 2016, the Mayo Clinic found a group of GFAP antibody-positive patients through retrospective detection of GFAP antibodies in the serum and cerebrospinal fluid of a group of patients. Therefore, a specific antibody targeting GFAP was redefined as a marker. According to A review of clinical cases found that the disease mainly involves the meninges, brain, spinal cord and optic nerve, and can be combined with tumors. It is a new type of meningoencephalomyelitis, and the disease is collectively named autoimmune glial fibrillary acidic protein astrocytes Autoimmune glial fibrillary acidic proteinastrocytopathy, referred to as autoimmune GFAP astrocytopathy. At present, the detection methods of glial f...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/6893G01N33/6854G01N2800/24G01N2333/47
Inventor 龙友明徐辉明刘天妮
Owner THE SECOND AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV
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