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Alpha-L-fucosidosidase active tested process and diagnostic reagent of alpha-L-fucosidosidase

A fucosidase and activity measurement technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of low value repeatability, low reaction sensitivity, negative measurement results, etc. The method is simple and convenient, the anti-interference is good, and the accuracy is high.

Active Publication Date: 2007-07-18
浙江伊利康生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method also has the following deficiencies: one, use the chromogenic substrate 2-chloro-p-nitrophenyl-alpha-L-fucoside (CNPF), which decomposes and turns yellow with the prolongation of time in the liquid state, The reagent blank is high, and the linearity of the reagent is affected by it. Second, the method is monitored at 405nm, which is easily interfered by bilirubin, fat turbidity and hemoglobin. The third is that the response sensitivity is not high, and the low-value repeatability is not ideal

Method used

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  • Alpha-L-fucosidosidase active tested process and diagnostic reagent of alpha-L-fucosidosidase
  • Alpha-L-fucosidosidase active tested process and diagnostic reagent of alpha-L-fucosidosidase
  • Alpha-L-fucosidosidase active tested process and diagnostic reagent of alpha-L-fucosidosidase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Reagent 1 (R1)

[0054] Citric acid buffer 100mmol / L (PH: 5)

[0055] Preservative NaN3 0.5mmol / L

[0056] Stabilizer EDTA.2Na 2mmol / L

[0057] Reagent 2 (R2)

[0058] Citric acid buffer 100mmol / L (PH: 4.9)

[0059] Substrate MPT-AFU 25mmol / L

[0060]Stabilizer EDTA.2Na 2mmol / L

[0061] Preservative NaN3 0.3mmol / L

Embodiment 2

[0063] Reagent 1 (R1)

[0064] Acetate buffer 50mmol / L (PH: 6.0)

[0065] Preservative NaN3 0.1mmol / L

[0066] Stabilizer EDTA.2Na 5mmol / L

[0067] Stabilizer NaCL 15mmol / L

[0068] Surfactant polyoxyethylene phenyl ether 0.1mmol / L

[0069] Reagent 2 (R2)

[0070] Acetate buffer 200mmol / L (PH: 4.2)

[0071] Substrate MPT-AFU 0.1mmol / L

[0072] Stabilizer EDTA.2Na 5mmol / L

[0073] Preservative NaN3 0.1mmol / L

[0074] Surfactant polyoxyethylene phenyl ether 0.1mmol / L

Embodiment 3

[0076] Reagent 1 (R1)

[0077] GOOd'S buffer 200mmol / L (PH: 5)

[0078] Preservative NaN3 5mmol / L

[0079] Stabilizer EDTA.2Na 0.1mmol / L

[0080] Surfactant Theist 5mmol / L

[0081] Reagent 2 (R2)

[0082] GOOd'S buffer 50mmol / L (PH: 4.9)

[0083] Substrate MPT-AFU 30mmol / L

[0084] Stabilizer EDTA.2Na 0.1mmol / L

[0085] Preservative NaN3 5mmol / L

[0086] Surfactant Theist 5mmol / L

[0087] Two, single reagent embodiment

[0088] Example 1

[0089] Phosphate buffer 100mmol / L (PH: 5.0)

[0090] Stabilizer EDTA.2Na 2mmol / L

[0091] Substrate MPT-AFU 45mmol / L

[0092] Preservative sorbitol 12mmol / L

[0093] Example 2

[0094] Citric acid buffer 50mmol / L (PH: 5.0)

[0095] Stabilizer iminodiacetic acid 5mmol / L

[0096] Substrate MPT-AFU 25mmol / L

[0097] Surfactant Theist 10mmol / L

[0098] Example 3

[0099] Acetate buffer 200mmol / L (PH: 4.2)

[0100] Stabilizer EDTA.2Na 0.1mmol / L

[0101] Substrate MPT-AFU 0.1mmol / L

[0102] Surfactant polyoxyethylene phenyl e...

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Abstract

The present invention relates to serum alpha-L-fucosidase activity measuring process and reagent, and the measuring process and reagent has short measuring period, high noise immunity and simple operation, and is suitable for various kinds of automatic biochemical analyzers. Of the technological scheme, the alpha-L-fucosidase activity measuring process includes adopting 6-methyl-2-thiopyridine-alpha-L-fucoside as substrate capable of producing 6-methyl-2-sulfhydyl pyridine under the action of alpha-L-fucosidase in the sample, and measuring the absorbance increasing rate of 6-methyl-2-sulfhydyl pyridine at 340 nm to find out the activity of alpha-L-fucosidase activity in the sample; and the alpha-L-fucosidase diagnosing reagent includes 6-methyl-2-sulfhydyl pyridine.

Description

technical field [0001] The invention relates to a method for measuring serum components and a reagent, in particular to a method for measuring serum α-L-fucosidase (AFU) activity and a reagent, and belongs to the technical field of biological preparations. Background technique [0002] α-L-fucosidase (α-L-fucosidase α-FU) is widely distributed in various tissues of the human body, such as liver, brain, kidney, pancreas and placenta, etc., exists in cells, is located in lysosomes, and is acidic Hydrolase, essentially a glycoprotein, participates in the catabolism of fucosyl-containing glycoproteins and glycolipids. In recent years, a large number of research data have shown that serum AFU is not only a marker of primary liver cancer and a reliable indicator for diagnosis, but also can be used for clinical monitoring of malignant transformation in patients with liver cirrhosis and observation indicators for diabetes control. [0003] The methods used to detect α-L-fucosidase ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/34
Inventor 王贤理蒙凯蔡其浩
Owner 浙江伊利康生物技术有限公司
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