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Alpha-hydroxybutyrate dehydrogenase detection kit and preparation method thereof

A technology of hydroxybutyrate dehydrogenase and detection kit, which is applied to the measurement of color/spectral characteristics, etc., and can solve the problem of backward quality control of α-hydroxybutyrate dehydrogenase determination, low accuracy of measurement results, and weak anti-interference To achieve the effect of speeding up clinical diagnosis, enhancing reaction sensitivity, and lower detection limit

Inactive Publication Date: 2015-03-25
CHONGQING ZHONGYUAN BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, the determination requires the enzyme and the substrate to act at a certain temperature for a period of time. During this period, the temperature may inactivate the enzyme. It is for these reasons that the quality control of the determination of α-hydroxybutyrate dehydrogenase is far behind. in other biochemical projects
For example, there are disadvantages such as expensive reagents, poor stability, low accuracy of measurement results, low sensitivity, weak anti-interference, cumbersome operation, and long time consumption.

Method used

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  • Alpha-hydroxybutyrate dehydrogenase detection kit and preparation method thereof
  • Alpha-hydroxybutyrate dehydrogenase detection kit and preparation method thereof
  • Alpha-hydroxybutyrate dehydrogenase detection kit and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] The kit of the present invention is exemplified as a double reagent, wherein:

[0041] Reagent R1:

[0042]

[0043] Reagent R2:

[0044]

Embodiment 2

[0045] Embodiment 2: Kit usage method.

[0046] 1. Reagent preparation, the reagent is a liquid double reagent, ready to use after opening the bottle, of which:

[0047] Reagent R1:

[0048]

[0049] Reagent R2:

[0050]

[0051] 2. Parameter setting of automatic biochemical analyzer:

[0052] (a) Detection wavelength: the main wavelength is 340nm, and the secondary wavelength is 405nm;

[0053] (b) Detection temperature: 37°C;

[0054] (c) Reaction time: 10 minutes, among which, the incubation time is 5 minutes, and the average absorbance change rate △A / min within 3 minutes is measured 2 minutes after adding reagent R2;

[0055] (d) Reaction direction: negative direction.

[0056] 3. Detection steps

[0057] (a) Mix 240ul reagent R1 with 6ul serum sample (to avoid hemolysis);

[0058] (b) Incubate the mixed solution at 37°C for 5 minutes;

[0059] (c) Add 60ul of reagent R2, react for 2 minutes and measure the average absorbance change rate ΔA / min within 3 minut...

Embodiment 3

[0071] Preparation and use of the comparison kit:

[0072] The biological buffer in reagent R1 is Tris buffer 50.0mmol / L, pH 7.0-7.5, and other reagents and experimental methods are the same as in Examples 1 and 2.

[0073]

[0074] Reagent R2:

[0075]

[0076] The performance test of the α-hydroxybutyrate dehydrogenase detection kit prepared in Example 3 is the same as in Example 2.

[0077] 1) Analytical sensitivity: Take a fixed-value α-hydroxybutyrate dehydrogenase sample with a concentration between 4 and 1200U / L to measure the change in absorbance, and repeat the measurement twice to get the average value. The results showed that its analytical sensitivity was 0.4581mA·L / U.

[0078] 2) Minimum detection limit: 5% BSA saline solution is used as a blank sample, and the blank sample should not contain the analyte. The detection was repeated 20 times continuously on the biochemical analyzer, and the detection results were recorded. The results showed that the lowe...

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Abstract

The invention discloses an alpha-hydroxybutyrate dehydrogenase detection kit. The alpha-hydroxybutyrate dehydrogenase detection kit comprises a reagent R1 and a reagent R2 which are independent to each other, wherein the reagent R1 is prepared from the following components including a biological buffering liquid 1, a metal ion complex, an alpha-hydroxybutyrate dehydrogenase reaction substrate, a surfactant 1 and a preservative 1; the reagent R2 is prepared from the following components including a biological buffering liquid 2, nicotinamide adenine dinucleotide reduced state, an activator, a surfactant 2 and a preservative 2. As adopting a mode of double reagents, the alpha-hydroxybutyrate dehydrogenase detection kit is high in detection sensitivity, low in detection limit, wide in linear test range, high in accuracy, good in repeatability, good in stability and strong in interference resistance, can be used for semi-automatic and fully automatic biochemical analyzers, is suitable for clinic popularization and application and in particular is capable of realizing rapid diagnosis of emergency treatment; the needed detection instruments (the biochemical analyzers) are generally used in various big hospitals and inspection centers.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to an alpha-hydroxybutyrate dehydrogenase detection kit and a preparation method thereof. Background technique [0002] α-hydroxybutyrate dehydrogenase (alpha-hydroxybutyrate dehydrogenase, a-HBDH) is an enzyme in the myocardial enzyme spectrum, which is ubiquitous in mammals. α-hydroxybutyrate dehydrogenase is widely distributed in various organs and tissues, mainly in myocardial red blood cells, white blood cells and kidneys. Like serum GOT, LDH, and creatine kinase (CK), it is widely present in brain cells. The activity of α-hydroxybutyrate dehydrogenase has a corresponding relationship with the change of LDH activity, which mainly represents the activity of isozymes rich in H subunits such as LDH1 and LDH2. Therefore, α-hydroxybutyrate dehydrogenase is commonly used to measure the activity of LDH1 in serum. The increase of α-hydroxybutyrate dehydrogenase activity is mainly...

Claims

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Application Information

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IPC IPC(8): G01N21/31
Inventor 周欢
Owner CHONGQING ZHONGYUAN BIOLOGICAL TECH
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