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Stable serum 5'-ribotide hydrolase detection reagent with strong anti-interference capability and detection method

A ribonucleotide and detection reagent technology, applied in the field of serum 5'-ribonucleotide hydrolase detection, can solve the problems of being susceptible to interference, unsuitable for automatic analysis, and many types of reagents

Active Publication Date: 2016-03-23
BIOBASE BIODUSTRY (SHANDONG) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the reagents of chemical colorimetry are easy to obtain, the operation is cumbersome and there are many kinds of reagents, so it is not suitable for automatic analysis and difficult to automate
Enzyme colorimetry, using multi-stage coupled enzymatic reactions, is simple to operate and suitable for automatic analysis, but has disadvantages such as poor stability and susceptibility to interference

Method used

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  • Stable serum 5'-ribotide hydrolase detection reagent with strong anti-interference capability and detection method
  • Stable serum 5'-ribotide hydrolase detection reagent with strong anti-interference capability and detection method
  • Stable serum 5'-ribotide hydrolase detection reagent with strong anti-interference capability and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Detection reagents for serum 5'-ribonucleotide hydrolase, including reagent R1 and reagent R2:

[0063] 1) The composition of its R1 is:

[0064] PIPES (1,4-piperazine diethanesulfonic acid) buffer (pH=7.6, 25°C) ···················· 100mmol / L,

[0065] 4-Aminoaminopyridine ························································································ 2.5mmol / L,

[0066] Peroxidase ································································································· 2KU / L,

[0067] nucleotide phosphorylase ························································································ 2KU / L,

[0068] xanthine oxidase ····························································································· 3KU / L,

[0069] Sodium β-Glycerophosphate ·························································································· 200mmol / L,

[0070] Bilirubin oxidase ······················································...

Embodiment 2

[0098] Interference test: Take fresh mixed serum, divide it into 2 equal parts, and then divide each equal part into 6 equal parts, add different interfering substances, so that the concentration in the serum can meet the requirements in Table 2. Then, the reagents obtained in Example 1 were used to compare and measure the activity of 5'-NT in the serum at the same time as the common and recognized serum 5'-ribonucleotide hydrolase (5'-NT) reagent in the market. The measurement results of each group after different interfering substances 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%.

[0099] As can be seen from Table 2, the reagent of embodiment 1 is tested at bilirubin≤20mg / dL, triglyceride≤1250mg / dL, hemoglobin≤500mg / dL, ascorbic acid≤220mg / dL, alkaline phosphatase≤1250U / L The results showed no significant interference. However, when the ...

Embodiment 3

[0103] Correlation experiment: use the formula of Example 1 to prepare reagents, and conduct a control test with the 5'-ribonucleotide hydrolase 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 , and the test results are shown in 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 0.9997, indicating that there is a great correlation between the two.

[0104] Table 3 Example 1 reagent and market common and recognized serum 5'-ribonucleotide hydrolase assay kit comparative detection results

[0105]

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Abstract

The invention relates to the technical field of serum 5'-ribotide hydrolase detection, in particular to a serum 5'-ribotide hydrolase detection reagent. A PIPES buffer solution is adopted, and various stabilizers are added to remarkably improve the stability of the reagent; beta-sodium glycerophosphate, bilirubin oxidase and ascorbic acid oxidase are added to effectively prevent interference caused by alkaline phosphatase, cholerythrin and ascorbic acid and greatly enhance the anti-interference capability of the reagent. Besides, a preferred novel amphoteric surfactant, dodecyl dimethyl betaine (BS-12), is added to prevent a reaction system from turbidity, enhance the stability of a substrate and improve the anti-interference capability of the reagent.

Description

technical field [0001] The invention relates to the technical field of serum 5'-ribonucleotide hydrolase detection, in particular to a serum 5'-ribonucleotide hydrolase detection reagent and a detection method using the detection reagent. Background technique [0002] 5'-ribonucleotide hydrolase (5'-NT) is a special hydrolase that can specifically hydrolyze inosine nucleotides into inosine and phosphate. This enzyme widely exists in human tissues, such as liver, gallbladder, intestine, brain, heart, pancreas, etc. 5'-NT enters bile through liver cell membrane, and then enters serum, which is one of the indicators for detecting liver and gallbladder diseases. This enzyme is significantly elevated in post-mastectomy patients with extrahepatic and extrahepatic bile duct obstruction and associated circulatory metastasis. 5'-NT is more sensitive than other enzymes in the liver for the diagnosis of hepatobiliary diseases. [0003] There are chemical colorimetric methods, rat...

Claims

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Application Information

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IPC IPC(8): C12Q1/48C12Q1/28C12Q1/26
CPCC12Q1/26C12Q1/28C12Q1/48
Inventor 赵新李志明谭柏清罗维晓王进
Owner BIOBASE BIODUSTRY (SHANDONG) CO LTD
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