Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method for measuring 5'-nucleotidase activity and Diagnostic reagent kit of 5'-nucleotidase

A diagnostic kit and nucleotidase technology, which is applied in the field of 5′-nucleotidase diagnostic kits to measure 5′-nucleotidase activity, and can solve problems such as poor accuracy, complicated methods, and isotope contamination. problems, to achieve the effect of good accuracy and accurate test results

Inactive Publication Date: 2006-03-22
王尔中
View PDF0 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The isotope substrate method is complicated and has isotope pollution, and requires an isotope analyzer, so it is difficult to practically popularize and apply
Inorganic phosphorus determination method is a method invented in 1925, the accuracy is not good, strong acid pollutes the environment, etc., it is not suitable for popularization and application

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0097] Embodiment one (single agent)

[0098] The 5'-nucleotidase diagnostic kit of the present embodiment comprises:

[0099] Buffer 80mmol / l

[0100] Adenosine monophosphate 1mmol / l

[0101] Pyruvate 1mmol / l

[0102] Pyruvate oxidase 5000U / l

[0103] Peroxidase 5000U / l

[0104] Stabilizer 50% (total volume)

[0105]4-amino antiarsenic 2mmol / l

[0106] Carbolic acid 10mmol / l

[0107] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, test main wavelength 495-505nm, test sub-wavelength above 600nm, the volume ratio of the tested 5'-nucleotidase sample to the reagent is 1 / 25. The reaction direction is positive reaction, the delay time is 1 minute, and the detection time is 2 minutes.

[0108] After the sample and reagents are added and allowed to mix, the following reactions occur:

[0109] Adenosine monophosphate + water 5 nucleotidase Adenosine + Phosphate

[0110] Phosphate + Pyruvate + Oxygen + Water pyruvate oxidase Ace...

Embodiment 2

[0116] Embodiment two (two doses)

[0117] The 5'-nucleotidase diagnostic reagent of the present embodiment has:

[0118] Reagent I

[0119] Buffer 100mmol / l

[0120] Pyruvate 5mmol / l

[0121] Pyruvate oxidase 12000U / l

[0122] Peroxidase 12000U / l

[0123] Stabilizer 50% (total volume)

[0124] 4-amino antiarsenic 1mmol / l

[0125] Reagent II

[0126] Buffer 100mmol / l

[0127] Adenosine monophosphate 3mmol / l

[0128] Stabilizer 30mmol / l

[0129] 2,4,6-tribromo-3-hydroxy-benzenesulfonic acid 1mmol / l

[0130] When measuring 5'-nucleotidase activity, the temperature is controlled at 30°C, the reaction time is 15 minutes, the main wavelength of the test is 546nm, the secondary wavelength of the test is above 630nm, and the volume ratio of the tested 5'-nucleotidase sample to the reagent is 1 / 25, the reaction direction is positive reaction, the delay time is 1 minute, and the detection time is 2 minutes.

[0131] The specific measurement steps are:

[0132] Adenosine m...

Embodiment 3

[0139] Embodiment three (three doses)

[0140] The 5′-nucleotidase diagnostic reagent of the present embodiment is three doses, including:

[0141] Reagent I

[0142] Buffer 120mmol / l

[0143] Pyruvate 10mmol / l

[0144] Stabilizer 50mmol / l

[0145] 3-methyl-2-benzothiazolone hydrazone 2mmol / l

[0146] Reagent II

[0147] Buffer 120mmol / l

[0148] Pyruvate oxidase 20000U / l

[0149] Peroxidase 20000U / l

[0150] Stabilizer 50% (total volume)

[0151] Dimethylaniline 2mmol / l

[0152] Reagent III

[0153] Buffer 120mmol / l

[0154] Adenosine monophosphate 5mmol / l

[0155] Stabilizer 50mmol / l

[0156] Set on the automatic biochemical analyzer: temperature 25°C, reaction time 20 minutes, test main wavelength 578nm, test sub-wavelength above 700nm, the volume ratio of the tested 5′-nucleotidase sample to the reagent is 1 / 25, The reaction direction is positive reaction, the delay time is 1 minute, and the detection time is 2 minutes.

[0157] The specific measurement step...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The present invention relates to 5í»-nucleotidase activity detecting method and 5í»-nucleotidase diagnosing reagent kit in medical detecting technology. The 5í»-nucleotidase diagnosing reagent kit includes buffering liquid, adenosine monophosphate, pyruvic acid, pyruvic oxidase, peroxidase, stabilizer, and reduction type chromogen combined together. Through mixing the sample and reagent in certain volume ratio to produce enzyme coupling reaction, and detecting in biochemical analyzer the main wavelength absorbency change speed, the activity of 5í»-nucleotidase is calculated. The present invention can obtain the measurement result in biochemical analyzer in high sensitivity, high precision and no contamination of various foreign and internal matters.

Description

technical field [0001] The invention relates to a method for measuring 5'-nucleotidase activity, and meanwhile, the invention also relates to a 5'-nucleotidase diagnostic kit for realizing the method, which belongs to the technical field of medical examination and determination. Background technique [0002] Medical research shows that the increase of 5'-nucleotidase (5NT) is mainly seen in obstructive jaundice, and can also be seen in liver cancer and hepatitis. In cholestasis complicated by cholangitis, primary and secondary biliary cirrhosis, and chronic hepatitis, the rate of 5NT elevation is higher than that of alkaline phosphatase; in liver tumors and hepatic granulomas, the sensitivity of 5NT elevation is higher than that of alkali sexual phosphatase. Because there is no physiological increase in 5'-nucleotidase activity, it is more sensitive and specific than alkaline phosphatase for the diagnosis of infantile liver disease and hepatic cholestasis of pregnancy. The...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/28
Inventor 王尔中
Owner 王尔中
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com