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Detection kit for activity of beta-glucocerebrosidase

A technology of glucocerebrosidase and detection kit, which is applied in the field of biochemical detection, can solve problems such as inability to effectively achieve early diagnosis, lack of rapid diagnostic reagent products, secondary injury to patients, etc., and achieve easy storage and transportation, blood demand The effect of small amount and accurate auxiliary diagnosis

Pending Publication Date: 2021-02-02
诺道中科(北京)生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there is no fast and convenient diagnostic reagent product for the clinical diagnosis of Gaucher disease in China. Usually, gene mutation detection, CT, X-ray and other means are combined with typical clinical symptoms such as facial changes, rough skin, joint deformation, and organ involvement. wait for diagnosis
However, for patients who lack typical clinical symptoms, none of the above methods can effectively achieve the purpose of early diagnosis, and there are drawbacks such as complicated operation, high cost, and repeated secondary injury to patients, and cannot be used as a more direct diagnostic standard. diagnosis of the disease

Method used

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  • Detection kit for activity of beta-glucocerebrosidase
  • Detection kit for activity of beta-glucocerebrosidase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] The composition of the detection kit of embodiment 1β-glucocerebrosidase activity

[0034] The detection kit comprises the following parts:

[0035] 1) Reaction solution: Weigh 10 mmol of 4-methylumbelliferyl-β-D-glucocerebroside, add it into purified water and stir to fully dissolve, adjust the pH to 4.0-6.0 with acetic acid-sodium acetate buffer solution, and constant volume to 1000mL.

[0036] 2) Precipitating agent: Weigh 0.25mol of barium hydroxide and 0.25mol of sodium carbonate into 1000mL of purified water and stir to fully dissolve.

[0037] 3) Stop solution: Weigh 0.5 mol of sodium carbonate and add it into 1000 mL of purified water, stir to dissolve fully, and adjust the pH of the solution to >10.0.

[0038] 4) Linear standard: Weigh 500nmol of 4-methylumbelliferone, add it to 1000mL of purified water and stir to fully dissolve; use this as the mother liquor, dilute to 40nmol / L, 100nmol / L, 200nmol / L, 400nmol with purified water / L, that's it.

[0039] 5) ...

Embodiment 2

[0040] The composition of the detection kit of embodiment 2β-glucocerebrosidase activity

[0041] The detection kit comprises the following parts:

[0042] 1) Reaction solution: Weigh 10 mmol of 4-methylumbelliferyl-β-D-glucocerebroside, add it into purified water and stir to fully dissolve, adjust the pH to 4.0-6.0 with PBS buffer, and dilute to 1000 mL.

[0043] 2) Precipitating agent: Weigh 0.2 mol of barium hydroxide and 0.3 mol of sodium carbonate into 1000 mL of purified water and stir to fully dissolve.

[0044] 3) Termination solution: Weigh 0.8 mol of sodium hydroxide and add it into 1000 mL of purified water, stir to fully dissolve, and adjust the pH of the solution to >10.0.

[0045] 4) Linear standard: Weigh 500nmol of 4-methylumbelliferone, add it to 1000mL of purified water and stir to fully dissolve; use this as the mother liquor, dilute to 40nmol / L, 100nmol / L, 200nmol / L, 400nmol with purified water / L, that's it.

[0046] 5) Reference substance: take at leas...

Embodiment 3

[0047] The composition of the detection kit of embodiment 3β-glucocerebrosidase activity

[0048] The detection kit comprises the following parts:

[0049] 1) Reaction solution: Weigh 10 mmol of 4-methylumbelliferyl-β-D-glucocerebroside, add it into purified water and stir to fully dissolve, adjust the pH with sodium citrate-dipotassium hydrogen phosphate buffer solution to 4.0- 6.0, set the volume to 1000mL.

[0050] 2) Precipitating agent: Weigh 0.3 mol of barium hydroxide and 0.2 mol of zinc sulfate into 1000 mL of purified water and stir to fully dissolve.

[0051] 3) Termination solution: Weigh 0.5 mol of sodium bicarbonate and 1 mol of sodium hydroxide, add them into 1000 mL of purified water, stir and fully dissolve, and adjust the pH of the solution to >10.0.

[0052] 4) Linear standard: Weigh 500nmol of 4-methylumbelliferone, add it to 1000mL of purified water and stir to fully dissolve; use this as the mother liquor, dilute to 40nmol / L, 100nmol / L, 200nmol / L, 400nmo...

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Abstract

The invention discloses a detection kit for the activity of beta-glucocerebrosidase, and belongs to the field of biochemical detection. The detection kit comprises a reaction solution, a precipitant,a stop buffer, a linear standard substance and a reference substance, wherein the linear standard substance is a 4-methylumbelliferone solution, and the reaction solution is a 4-methylumbelliferyl-beta-D-glucocerebroside solution. The kit can rapidly and accurately perform in vitro detection on the activity of the beta-glucocerebrosidase in a human blood sample, and the goal of rapidly, conveniently and accurately assisting in diagnosing gaucher disease patients in batches can be achieved by detecting collected filter paper dried blood spot sample or venous blood samples; and the kit has highdetection sensitivity, good selectivity, high anti-interference capability and good reproducibility, is simple and convenient to operate, has high product stability, and is easy to use clinically.

Description

technical field [0001] The invention relates to the field of biochemical detection, in particular to a detection kit for β-glucocerebrosidase activity. Background technique [0002] Gaucher disease (GD), namely glucocerebrosidosis, is a familial disorder of glucose and lipid metabolism. It is a chromosomal recessive inheritance and is the most common type of lysosomal deposition disease. Glucocerebroside is a soluble glycolipid substance, which is one of the components of cells and widely exists in the human body. Due to the lack of glucocerebrosidase, glucocerebroside accumulates in the mononuclear-macrophages of the liver, spleen, bone and central nervous system, resulting in corresponding clinical manifestations. Gaucher disease occurs all over the world. The Jewish population has a higher incidence. Since the first report in my country in 1948, there have been reports from all over the country, especially in Hebei, Shandong, Henan and Liaoning. [0003] Gaucher disea...

Claims

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Application Information

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IPC IPC(8): C12Q1/34
CPCC12Q1/34G01N2333/924C12Q2334/22
Inventor 王永凯马蒙蒙唐赞王瑞刘立明卢孟孟孙杰胖铁良李建中
Owner 诺道中科(北京)生物科技有限公司
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