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A kit for enzymatic detection of d-3-hydroxybutyric acid and its preparation method

A D-3-, reagent kit technology, applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of bilirubin interference, impact detection, negative value, etc., and achieve anti-bilirubin interference Enhanced performance, great clinical application value, and improved sensitivity

Active Publication Date: 2020-04-10
SHANGHAI FOSUN LONG MARCH MEDICAL SCI CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method has low sensitivity and poor anti-interference ability, especially bilirubin has serious interference on the detection, and negative values ​​often appear, which affects the detection. The composition of the existing enzymatic kit: reagent R1Tris buffer 100mmol / L oxalic acid 20mmol / Lβ -Hydroxybutyrate dehydrogenase 1KU / L Reagent R2INT 1mmol / L NAD+2.5mmol / L

Method used

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  • A kit for enzymatic detection of d-3-hydroxybutyric acid and its preparation method
  • A kit for enzymatic detection of d-3-hydroxybutyric acid and its preparation method
  • A kit for enzymatic detection of d-3-hydroxybutyric acid and its preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Preparation of D-3-hydroxybutyric acid detection kit

[0029] composition:

[0030] Reagent 1: (3L) (reagent volume)

[0031]

[0032] Reagent 2: (1L) (reagent volume)

[0033]

[0034]

[0035] preparation:

[0036] (1) Preparation of reagent 1: (3L)

[0037] (1) First add 2.4L of deionized water to the container;

[0038] (2) Tris buffer solution, lauryl polyethylene glycol ether, sodium azide are added successively;

[0039] (3) Add enzyme: D-3-hydroxybutyrate dehydrogenase

[0040] (4) Finally, add deionized water until the total volume is 3L and mix evenly to obtain;

[0041] (2) Preparation of reagent 2: (1L)

[0042] Add oxalate, 3-acetylpyridine oxidized coenzyme, and sodium azide deionized water to the container in sequence until the total volume is 1 L, and mix well.

Embodiment 2

[0044] Reagent 1: (3L) (reagent volume)

[0045]

[0046] Reagent 2: (1L) (reagent volume)

[0047]

[0048]

[0049] The preparation method is the same as in Example 1.

Embodiment 3

[0051] Reagent 1: (3L) (reagent volume)

[0052]

[0053] Reagent 2: (1L) (reagent volume)

[0054]

[0055] Preparation method is the same as embodiment 1

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Abstract

The invention provides a kit of detecting D-3-hydroxybutyric acid through an enzymic method. The kit is composed of a first reagent and a second reagent according to the volume part ratio of 3:1. 3L of the first reagent includes 10-200 mmol / L of a buffer liquid, 50-200 U / L of a D-3-hydroxybutyric dehydrogenase, 1-3 g / L of Brij35 (dodecyl polyglycol ether), 0.1-2 g / L of sodium azide, and deionized water being added to the components until the total volume is 3L; and 1L of the second reagent includes: 10-200 mmol / L of oxalate, 0.1-10 mmol / L of 3-acetylpyridine nicotinamide adenine dinucleotide, 0.1-2 g / L of sodium azide, and deionized water being added to the components until the total volume is 1L. The kit has strong performance of resisting bilirubin interference, has high reagent sensitivity, is improved in detection accuracy and has great clinical application value.

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] Diabetic ketoacidosis is a common clinical disease, frequently-occurring disease, caused by increased blood ketones, D-3-hydroxybutyric acid is the main component of ketones in the blood (accounting for 78%), it can reflect the ketones in the blood generated situation. 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 . Because in ketoacidosis, the patient has an antagonistic effect on insulin, resulting in protein synthesis disorder and protein degradation metabolism, therefore, by measuring the fasting blood sugar in the morning, it ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/32
Inventor 景晟孙卫兵张跃建
Owner SHANGHAI FOSUN LONG MARCH MEDICAL SCI CO LTD
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