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Kit for determination of alpha-hydroxybutyrate dehydrogenase and preparation method thereof

A hydroxybutyrate dehydrogenase and kit technology are applied in the fields of medicine and biochemistry, which can solve the problems of inaccurate measurement results of alpha-hydroxybutyrate dehydrogenase, and achieve the effects of improving stability and improving stability.

Inactive Publication Date: 2016-11-09
ANHUI IPROCOM BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved by the present invention is to provide a test kit for measuring α-hydroxybutyrate dehydrogenase and its preparation method in order to overcome the inaccurate defect of the existing α-hydroxybutyrate dehydrogenase assay results

Method used

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  • Kit for determination of alpha-hydroxybutyrate dehydrogenase and preparation method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] The kit of the present invention includes reagent R1 and reagent R2 two-liquid components independent of each other, wherein

[0061] Reagent R1:

[0062] Tris Buffer 180mmol / L

[0063] α-ketobutyric acid 5mmol / L

[0064] Disodium EDTA 1.0g / L

[0065] Sodium azide 0.20g / L

[0066] Its solvent is purified water.

[0067] Reagent R2:

[0068] Tris Buffer 180mmol / L

[0069] NADH 2.0mmol / L

[0070] Sodium azide 0.20g / L

[0071] Its solvent is purified water.

Embodiment 2

[0073] The kit of the present invention includes reagent R1 and reagent R2 two-liquid components independent of each other, wherein

[0074] Reagent R1:

[0075] 3-Morpholine propanesulfonic acid buffer 350mmol / L

[0076] α-ketobutyric acid 10mmol / L

[0077] Disodium EDTA 2g / L

[0078] Sodium azide 0.01g / L

[0079] Its solvent is purified water.

[0080] Reagent R2:

[0081] 3-Morpholine propanesulfonic acid buffer 350mmol / L

[0082] NADH 4.0mmol / L

[0083] Sodium azide 0.01g / L

[0084] Its solvent is purified water.

Embodiment 3

[0086] Kit preparation and method of use

[0087] 1. Prepare the reagent according to the content of the following components:

[0088] Reagent R1:

[0089] Tris Buffer 180mmol / L

[0090] α-ketobutyric acid 5mmol / L

[0091] Disodium EDTA 1.0g / L

[0092] Sodium azide 0.20g / L

[0093] Its solvent is purified water.

[0094] Reagent R2:

[0095] Tris Buffer 180mmol / L

[0096] NADH 2.0mmol / L

[0097] Sodium azide 0.20g / L

[0098] Its solvent is purified water.

[0099] 2. Parameter setting of automatic biochemical analyzer

[0100] (a) Detection temperature: 37°C;

[0101] (b) Detection wavelength: main wavelength 340nm, secondary wavelength 405nm;

[0102] (c) Reaction time: 8 minutes, among them, the incubation time is 5 minutes, measure and read the absorbance A1 immediately after adding the reagent R2, read the absorbance A2 after 3 minutes, and calculate the change of absorbance ΔA = A2-A1;

[0103] (d) Reaction direction: negative reaction.

[0104] 3. Detectio...

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Abstract

The invention discloses a kit for determination of alpha-hydroxybutyrate dehydrogenase and a preparation method thereof, wherein the kit includes double-liquid components of a reagent R1 and a reagent R2 which are independent with each other and includes the components and the corresponding content: the reagent R1 including 1-350 mmol / L of a buffer liquid, 0.01-10 mmol / L of alpha-ketone butyric acid, 0.1-2 g / L of disodium ethylenediamine tetraacetate, 0.01-0.40 g / L of a stabilizing agent, and a solvent being purified water; and the reagent R2 including 1-350 mmol / L of a buffer liquid, 0.1-4.0 mmol / L of NADH, 0.01-0.40 g / L of a stabilizer, and a solvent being purified water. The preparation method includes the steps: according to the following component content, preparing the reagents; mixing a to-be-tested sample with the reagent R1 and the reagent R2, and carrying out full reaction; determining the absorbance difference value after the reaction with a fully automatic biochemical analyzer; and calculating the concentration of the alpha-hydroxybutyrate dehydrogenase in the sample according to the absorbance change value. The kit has the advantages of high accuracy, good precision and the like.

Description

technical field [0001] The invention relates to the technical fields of medicine and biochemistry, in particular to a kit for measuring α-hydroxybutyrate dehydrogenase and a preparation method thereof. Background technique [0002] α-Hydroxybutyrate dehydrogenase (α-HBDH) is not an independent specific enzyme, but a general term for lactate dehydrogenase isozymes LDH1 and LDH2 containing H subunits. Determination of α-hydroxybutyrate dehydrogenase, which actually reflects the activity of lactate dehydrogenase isoenzymes LDH1 and LDH2, is of great significance for the diagnosis of myocardial disease and liver disease. The activity of α-HBDH is consistent with that of LDH, but it can better reflect the change of LDH1 activity, and its specificity is higher than that of total LDH activity. Combined with LDH, AST, CK, and CK-MB to form myocardial enzyme spectrum, it is more meaningful for the diagnosis of myocardial infarction. [0003] There are many current detection methods...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/32
CPCC12Q1/32G01N2333/904
Inventor 蔡晓辉庄庆华吴铮徐运
Owner ANHUI IPROCOM BIOTECH CO LTD
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