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Detection kit for alpha-galactosidase activity

A technology of galactosidase and detection kit, which is applied in the field of biochemical detection to achieve the effects of high sensitivity, good reproducibility and good stability

Pending Publication Date: 2021-01-05
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] The object of the present invention is to provide a detection kit for α-galactosidase activity to solve the problems in the prior art above. Glycosidase activity, auxiliary diagnosis of Fabry disease patients, has the characteristics of trace, fast and convenient, and solves the problem that there is no similar mature detection kit product currently used

Method used

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  • Detection kit for alpha-galactosidase activity

Examples

Experimental program
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Effect test

Embodiment 1

[0032] The composition of the detection kit of embodiment 1α-galactosidase activity

[0033] The detection kit comprises the following parts:

[0034] 1) Reaction solution: Weigh 20mmol of 4-methylumbelliferyl-α-D-galactoside, add it into purified water and stir to fully dissolve, adjust the pH to 4.0-6.0 with acetic acid-sodium acetate buffer, and set the volume to 1000mL .

[0035] 2) Precipitating agent: Weigh 0.125mol of barium hydroxide and 0.125mol of sodium carbonate into 1000mL of purified water and stir to fully dissolve.

[0036] 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.

[0037] 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.

[0038] 5) Reference substanc...

Embodiment 2

[0039]The composition of the detection kit of embodiment 2α-galactosidase activity

[0040] The detection kit comprises the following parts:

[0041] 1) Reaction solution: Weigh 20 mmol of 4-methylumbelliferyl-α-D-galactoside, add it into purified water, stir and fully dissolve, adjust the pH to 4.0-6.0 with PBS buffer, and dilute to 1000 mL.

[0042] 2) Precipitating agent: Weigh 0.1 mol of barium hydroxide and 0.15 mol of sodium carbonate into 1000 mL of purified water and stir to fully dissolve.

[0043] 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.

[0044] 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.

[0045] 5) Reference substance: take at least 3 whole bl...

Embodiment 3

[0046] The composition of the detection kit of embodiment 3α-galactosidase activity

[0047] The detection kit comprises the following parts:

[0048] 1) Reaction solution: Weigh 20 mmol of 4-methylumbelliferyl-α-D-galactoside, add it into purified water and stir to fully dissolve it, adjust the pH to 4.0-6.0 with sodium citrate-dipotassium hydrogen phosphate buffer solution, Dilute to 1000mL.

[0049] 2) Precipitating agent: Weigh 0.15 mol of barium hydroxide and 0.1 mol of zinc sulfate and add them into 1000 mL of purified water and stir to fully dissolve.

[0050] 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.

[0051] 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 ...

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Abstract

The invention discloses a detection kit for alpha-galactosidase activity, and belongs to the field of biochemical detection. The detection kit comprises a reaction solution, a precipitant, a stop solution, 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-methylumbelliferone acyl-alpha-D-galactoside solution. The kit can rapidly and accurately carry out in-vitro detection on the activity of alpha-galactosidase in a human blood sample, and can achieve the purpose of rapidly, conveniently and accurately assisting in diagnosing Fabry disease patients in batches by detecting a collected filter paper dry blood slice sample or a venous blood sample; and the kit is high in detection sensitivity, good in selectivity, strong in anti-interference capability, good in reproducibility, simple and convenient to operate, good in product stability and easy to use clinically.

Description

technical field [0001] The invention relates to the field of biochemical detection, in particular to a detection kit for alpha-galactosidase activity. Background technique [0002] Fabry disease is an X-chromosome hereditary multisystem lysosomal storage disease caused by mutations in the α-galactosidase A (α-Gal A) gene. The incidence rate of male newborns is 1 / 110000~1 / 40000. Both men and women are affected, and the onset is mostly from childhood to adolescence, and women's symptoms are usually milder than men's. Fabry disease is due to the mutation of the gene encoding α-galactosidase A, which leads to the partial or complete loss of the function of α-galactosidase A encoded by it, and the normal degradation of trimeric hexosylceramide is blocked, and the undegraded substrate Substances accumulate in the cell lysosomes of various tissues, causing dysfunction of related tissues. Fabry disease is divided into two types: classic and atypical: classic patients with early o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/34
CPCC12Q1/34C12Q2334/22G01N2333/94
Inventor 王瑞卢孟孟唐赞孙杰胖铁良李建中
Owner 诺道中科(北京)生物科技有限公司
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