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Stable and high-interference resistance serum adenosine deaminase detection reagent and detection method

A technology of serum adenosine deaminase and detection reagent, applied in the field of serum adenosine deaminase detection, can solve the problems of difficult to control electrode response signal and response time, poor repeatability and accuracy, uncomfortable measurement, etc. Anti-interference ability, avoid interference, prevent turbid effect

Inactive Publication Date: 2016-11-16
BIOBASE BIODUSTRY (SHANDONG) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the determination methods of adenosine deaminase include Bowers chromogenic method, ammonia gas-sensing electrode method, enzymatic method, enzyme coupling method, etc.; The influence of ammonium ions; although the ammonia gas sensitive electrode method has strong anti-interference ability, it is difficult to control the response signal and response time of the electrode, and is not suitable for the determination of large quantities of clinical samples; the enzymatic method is more sensitive and can realize automatic analysis, but The repeatability and accuracy are poor; the results of the enzyme coupling method are accurate, simple and practical, but are easily interfered by substances such as ascorbic acid and bilirubin

Method used

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  • Stable and high-interference resistance serum adenosine deaminase detection reagent and detection method
  • Stable and high-interference resistance serum adenosine deaminase detection reagent and detection method
  • Stable and high-interference resistance serum adenosine deaminase detection reagent and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Detection reagents for serum adenosine deaminase, including reagent R1 and reagent R2:

[0057] 1) The composition of its R1 is:

[0058] HEPES (4-hydroxyethylpiperazineethanesulfonic acid) buffer solution (pH=8.0, 25°C) ·· 100mmol / L,

[0059] 4-Aminoaminopyridine ································· 5mmol / L,

[0060] PNP (purine nucleoside phosphorylase) ·························· 1.5KU / L,

[0061] XOD (Xanthine Oxidase) ······························ 3.1KU / L,

[0062] POD (Peroxidase) ································ 5KU / L,

[0063] Bilirubin oxidase ··································· 3KU / L,

[0064] ascorbate oxidase ································· 3.5KU / L,

[0065]Glycerol ········································· 4ml / L,

[0066] polyethylene glycol 6000 ··································· 5g / L,

[0067] ethylene glycol ········································· 6ml / L,

[0068] Mannitol ········································· 25g / L,

[0069] Tre...

Embodiment 2

[0088] Interference test: Take fresh mixed serum, divide it into 2 equal parts, then divide each equal part into 5 equal parts, add different interfering substances, so that the concentration in the serum reaches Figure 4 Requirements; Then respectively use the reagent obtained in embodiment 1, and the serum adenosine deaminase (ADA) reagent that is common and approved in the market is compared and measured the activity of ADA in the serum simultaneously, the measured result of the control group is different from that of each group after adding different interfering substances. See the test results Figure 4 ; Relative deviation (%) = (measuring mean value of interference sample - measuring mean value of control sample) / measuring mean value of control sample × 100%;

[0089] Depend on Figure 4 It can be seen that the reagent of Example 1 has no obvious interference to the test results when total bilirubin≤30mg / dL, triglyceride≤3000mg / dL, hemoglobin≤800mg / dL, ascorbic acid...

Embodiment 3

[0091] Correlation experiment: using the formula in Example 1 to prepare the reagents, and conduct a control test with the adenosine deaminase (ADA) kit of a company approved by the State Food and Drug Administration, which is common in the market, and test 20 clinical serum samples at the same time. Test results such as Figure 5 As shown, and the correlation curves of the two reagents were obtained (as figure 1 As shown), the test results show that the correlation coefficient of the two kits is 0.9996, which shows that there is a great correlation between the two.

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Abstract

The invention relates to the technical field of detection of serum adenosine deaminase and in particular relates to a serum adenosine deaminase detection reagent. R1 comprises buffer solution, 4-aminoantipyrine, PNP, XOD, POD, bilirubin oxidase, ascorbic acid oxidase, glycerin, polyethylene glycol 6000, ethylene glycol, mannitol, mycose, BSA, alkyl glycoside and preservatives; R2 comprises buffer solution, adenosine, 3-hydroxy-2,4,6-triiodobenzoic acid, glycerin, polyethylene glycol 6000, ethylene glycol, mannitol, mycose, BSA, alkyl glycoside and preservatives. With the adoption of the novel Trinder reaction chromogen substance 3-hydroxy-2,4,6-triiodobenzoic acid, multiple stabilizers are added, and the stability of the reagent is obviously improved; and due to the added bilirubin oxidase, ascorbic acid oxidase and novel nonionic surfactant alkyl glycoside (APG), interference of bilirubin and ascorbic acid is avoided, a turbid reaction system is prevented, and the substrate stability and the interference resistance of the reagent are enhanced.

Description

technical field [0001] The invention relates to the technical field of serum adenosine deaminase detection, in particular to a serum adenosine deaminase detection reagent and a detection method using the detection reagent. Background technique [0002] Adenosine deaminase (ADA) is an important enzyme in the metabolism of purine nucleotides in the human body. It is widely distributed in various tissues of the human body, among which the thymus, spleen and other lymphoid tissues have the highest content, while the liver, lung, kidney and The content of skeletal muscle is low; ADA in serum mainly comes from the liver, which is a sensitive indicator reflecting liver damage and can be used as one of the routine inspection items of liver function. The increase or decrease of the enzyme activity reflects the degree of liver cell damage and recovery; chronic The ADA activity of erythrocytes in patients with hemolysis was significantly increased; the ADA activity in serum and tissue ...

Claims

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Application Information

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IPC IPC(8): C12Q1/48C12Q1/34C12Q1/26C12Q1/28
CPCC12Q1/48C12Q1/26C12Q1/28C12Q1/34C12Q2326/00C12Q2326/96
Inventor 李志明甘宜梧谭柏清王珍李贵雨
Owner BIOBASE BIODUSTRY (SHANDONG) CO LTD
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