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D-3-hydroxybutyrate enzyme detection kit and preparation method thereof

A D-3-, kit technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of long capillary isotachyphoresis operation time, unacceptable chemical substances, difficult to use in large quantities, etc. Great clinical application value, improved stability, strong protective effect

Active Publication Date: 2016-08-17
SHANGHAI FOSUN PHARMA (GROUP) CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Radiochemical methods are not suitable for clinical application due to radiation, chemical substances are difficult to accept, capillary isotachophoresis takes a long time, and the operation is complicated.
Enzymatic reagents cannot be stored for a long time after being made into liquid reagents, and the price of enzymes is very expensive and difficult to use in large quantities. Therefore, it is necessary to improve the determination method

Method used

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  • D-3-hydroxybutyrate enzyme detection kit and preparation method thereof
  • D-3-hydroxybutyrate enzyme detection kit and preparation method thereof
  • D-3-hydroxybutyrate enzyme detection kit and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 Preparation of D-3-hydroxybutyric acid detection kit

[0035] composition:

[0036] Reagent 1: (3L) (reagent volume)

[0037]

[0038] Reagent 2: (1L) (reagent volume)

[0039]

[0040] preparation:

[0041] (1) Preparation of reagent 1: (3L)

[0042] (1) First add 2.4L of deionized water to the container;

[0043] (2) Add tris buffer, polyoxypropylene polyoxyethylene copolymer (F108), and sodium azide in sequence;

[0044] (3) Add enzyme: D-3-hydroxybutyrate dehydrogenase

[0045] (4) Finally, add deionized water to a total volume of 3L and mix evenly to obtain;

[0046] (2) Preparation of reagent 2: (1L)

[0047] Add oxalate, oxidized coenzyme, and sodium azide deionized water to the container in sequence until the total volume is 1 L, and mix well.

Embodiment 2

[0049] Reagent 1: (3L) (reagent volume)

[0050]

[0051] Reagent 2: (1L) (reagent volume)

[0052] Oxalate 3.5g

[0053] Oxidized Coenzyme 3g

[0054] Sodium azide 1g

[0055] Deionized water was added to a total volume of 1 L.

[0056] The preparation method is the same as in Example 1.

Embodiment 3

[0058] Reagent 1: (3L) (reagent volume)

[0059]

[0060] Reagent 2: (1L) (reagent volume)

[0061] Oxalate 5g

[0062] Oxidized Coenzyme 3g

[0063] Sodium azide 1g

[0064] Deionized water was added to a total volume of 1 L.

[0065] Preparation method is the same as embodiment 1

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Abstract

The invention provides an enzymatic method detection kit of D-3-hydroxybutyric acid. The kit comprises a reagent 1 and a reagent 2 according to a ratio of 3: 1. The reagent 1 (with the volume of 3L) comprises 10-200mmol / L of a buffer solution, 50-200U / L of D-3-hydroxybutyric acid dehydrogenase, 1-3g / L of a polyoxypropylene-polyoxyethylene copolymer, 0.1-2g / L of sodium azide and the balance deionized water. The reagent 2 (with the volume of 1L) comprises 10-200mmol / L of oxalate, 0.1-10mmol / L of oxidized coenzyme, 0.1-2g / L of sodium azide and the balance deionized water. The used D-3-hydroxybutyric acid dehydrogenase stabilizing agent can good protect enzyme in the reagent so that the D-3-hydroxybutyric acid kit stability is improved, a cost is greatly reduced and a large clinical application value is obtained.

Description

technical field [0001] The invention relates to biological reagents, in particular to a detection kit, in particular to a kit for enzymatic detection of D-3-hydroxybutyric acid and a preparation method thereof. Background technique [0002] D-3-hydroxybutyric acid is the main component of ketone bodies in blood (accounting for 78%), which can reflect the production of ketone bodies. When ketoacidosis occurs, D-3-hydroxybutyric acid is significantly increased. Therefore, D-3-hydroxybutyric acid can be used for early diagnosis and treatment monitoring of ketoacidosis, and can guide the treatment and efficacy monitoring of ketoacidosis . In ketoacidosis, the patient has an antagonistic effect on insulin, resulting in protein synthesis disorders and protein metabolism. This can be used to measure the body fat and protein in patients with severe injuries, acute illnesses, sepsis and nutritional support by measuring morning fasting blood sugar. mobilization situation. For weigh...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/32
CPCC12Q1/32
Inventor 景晟孙卫兵张跃建
Owner SHANGHAI FOSUN PHARMA (GROUP) CO LTD
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