Stable alpha-hydroxybutyrate dehydrogenase reagent with high interference resistance capacity and detection method

A hydroxybutyrate dehydrogenase and detection method technology, which is applied in the field of α-hydroxybutyrate dehydrogenase detection reagents, can solve the problems of poor stability of the reagent substrate NADH, etc., and achieve enhanced stability and anti-interference ability, Inexpensive and easy to use

Active Publication Date: 2015-10-21
BIOBASE BIODUSTRY (SHANDONG) CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This method is simple and fast, and is suitable for automatic biochemical analysis, but the ...

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  • Stable alpha-hydroxybutyrate dehydrogenase reagent with high interference resistance capacity and detection method
  • Stable alpha-hydroxybutyrate dehydrogenase reagent with high interference resistance capacity and detection method
  • Stable alpha-hydroxybutyrate dehydrogenase reagent with high interference resistance capacity and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] α-Hydroxybutyrate dehydrogenase detection reagent, including reagent R1 and reagent R2:

[0040] 1) The composition of its R1 is:

[0041] PIPES (piperazine-1,4-diethanesulfonic acid) buffer (pH=7.4, 25°C) ·························· 50mmol / L

[0042] Disodium ethylenediaminetetraacetic acid (EDTA-2Na) ····································································· 3mmol / L,

[0043] alpha-ketobutyric acid ··································································································· 5mmol / L,

[0044] Lauryl dimethyl betaine ·············································································· 2g / L,

[0045] Preservative NaN 3······························································································· 0.5g / L

[0046] 2) The components of reagent R2 are:

[0047] PIPES (piperazine-1,4-diethanesulfonic acid) buffer (pH=7.4, 25°C) ·························· 100mmol / L

[0048] NADH ····························...

Embodiment 2

[0061] Interfering test

[0062] Take fresh mixed serum, divide it into 2 equal parts, and then divide each equal part into 4 equal parts, add different interfering substances, so that the concentration in the serum reaches the requirements in Table 2. Then respectively use the reagent obtained in Example 1, and the α-hydroxybutyrate dehydrogenase (α-HBDH) reagent that is common and recognized in the market to compare and measure the activity of α-HBDH in the serum simultaneously, and the measured results of the control group are the same as those after adding different interfering substances. The measurement results of each group are shown in Table 2. Relative deviation (%) = (measuring mean value of interference samples - measuring mean value of control samples) / measured mean value of control samples × 100%.

[0063] It can be seen from Table 2 that the reagent of Example 1 has no obvious interference on the test results when ascorbic acid≤50mg / dL, bilirubin≤1026μmol / L, and...

Embodiment 3

[0067] correlation experiment

[0068] Utilize the formula of Example 1 to prepare the reagents, and carry out comparative detection with the α-hydroxybutyrate dehydrogenase kit of a company approved by the State Food and Drug Administration, which is common in the market, and detect 20 clinical serum samples at the same time, and the detection results are shown in the table 3. And obtained the correlation curve of the two reagents (such as figure 1 Shown), the test results show that the correlation coefficient of the two kits is 0.9994, indicating that there is a great correlation between the two.

[0069] Table 3 Comparative test results of the reagent in Example 1 and the common and recognized α-hydroxybutyrate dehydrogenase assay kit in the market

[0070] .

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Abstract

The invention relates to the technical field of the detection of alpha-hydroxybutyrate dehydrogenase, and particularly to an alpha-hydroxybutyrate dehydrogenase reagent. The reagent R1 is prepared from a buffering solution, EDTA-2Na, alpha-ketobutyric acid, dodecyl dimethyl betaine and preservative; the reagent R2 is prepared from a buffering solution, NADH, BSA, saccharose, trehalose, mannitol, dodecyl dimethyl betaine and preservative. By adopting the PIPES buffering solution and adding the stabilizer BSA, the saccharose, the trehalose and the mannitol, the stability of the reagent is greatly improved; by adopting the ampholytic surfactant dodecyl dimethyl betaine, not only is the determination performance remarkably improved, but also the stability and interference resistance capacity of the reagent are improved.

Description

technical field [0001] The invention relates to the technical field of alpha-hydroxybutyrate dehydrogenase detection, in particular to a detection reagent for alpha-hydroxybutyrate dehydrogenase and a detection method using the detection reagent. Background technique [0002] α-Hydroxybutyrate dehydrogenase (α-HBDH), one of the enzymes in the myocardial zymogram, is essentially an LDH1-type isoenzyme, mainly derived from the myocardium, kidney and red blood cells, and is most abundant in the myocardium. The determination of α-hydroxybutyrate dehydrogenase activity is commonly used in the auxiliary diagnosis of acute myocardial infarction (AMI), which provides a powerful quantitative index for the clinical diagnosis, condition observation, curative effect analysis and prognosis estimation of AMI. Its determination is more specific and valuable for the diagnosis of AMI than LDH. In addition, the increase of α-hydroxybutyrate dehydrogenase is mainly seen in myocardial infarcti...

Claims

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Application Information

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IPC IPC(8): C12Q1/32
Inventor 甘宜梧李志明谭柏清李静王绮
Owner BIOBASE BIODUSTRY (SHANDONG) CO LTD
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