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Detection kit for activity of alpha-L-iduronidase

A technology of idurosidase and detection kit, which is applied in the field of medical diagnosis, can solve the problems of ineffective early diagnosis, secondary injury of patients, complicated operation, etc., and achieves easy storage and transportation, simple and easy operation, and selective Good results

Pending Publication Date: 2020-11-06
北京中科医学检验实验室有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

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 alpha-L-iduronidase
  • Detection kit for activity of alpha-L-iduronidase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] The composition of the detection kit of embodiment 1α-L-iduronidase activity

[0034] The detection kit comprises the following parts:

[0035] 1) Reaction solution: Weigh 10 mmol of 4-methylumbelliferyl-β-D-galactopyranoside, 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 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) Reference subst...

Embodiment 2

[0040] The composition of the detection kit of embodiment 2α-L-iduronidase activity

[0041] The detection kit comprises the following parts:

[0042] 1) Reaction solution: Weigh 10 mmol of 4-methylumbelliferyl-β-D-galactopyranoside, 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.

[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 least 3 wh...

Embodiment 3

[0047] The composition of the detection kit of embodiment 3α-L-iduronidase activity

[0048] The detection kit comprises the following parts:

[0049] 1) Reaction solution: Weigh 10 mmol of 4-methylumbelliferyl-β-D-galactopyranoside, add it into purified water and stir to fully dissolve it, and adjust the pH 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, 400nmol with purified water / L, that's it.

[0053] 5) Reference sub...

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Abstract

The invention discloses a detection kit for the activity of alpha-L-iduronidase, and belongs to the field of medical diagnosis. The detection kit comprises a reaction solution, a precipitator, a stopsolution, 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-beta-D-galactopyranoside solution. The kit can be used for rapidly and accurately detecting the activity of the alpha-L-iduronidase in a human blood sample in vitro; by detecting a collected filter paper dried blood spot sample or venous blood sample, the purpose of rapidly, conveniently and accurately assisting in diagnosis of mucopolysaccharidosis type I patients in batches can be achieved; 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 medical diagnosis, in particular to a detection kit for α-L-iduronidase activity. Background technique [0002] Mucopolysaccharide metabolism disorder is an X-linked recessive genetic disease that occurs due to the mutation of the hydrolase that degrades mucopolysaccharides in the lysosome of human cells, resulting in the loss of its activity, mucopolysaccharides cannot be degraded and metabolized, and eventually accumulate in the body. The disease is a very important type of lysosomal storage disease, which can be divided into 7 types including types Ⅰ, Ⅱ, Ⅲ, Ⅳ, Ⅵ, Ⅶ, and Ⅸ. Although the pathogenic genes and clinical manifestations of each type are different, But because the storage substrates are all mucopolysaccharides, they are collectively referred to as mucopolysaccharide metabolism disorders. [0003] Mucopolysaccharide is a long-chain complex sugar molecule, which is formed by connecting disaccharide units...

Claims

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Application Information

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IPC IPC(8): C12Q1/34
CPCC12Q1/34G01N2800/042
Inventor 刘立明齐晓丽唐赞王瑞卢孟孟马蒙蒙王永凯胖铁良李建中
Owner 北京中科医学检验实验室有限公司
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