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Method for testing activity of 5'-nucleotidase, diagnosis kit for 5'-nucleotidase

A diagnostic kit and nucleotidase technology, applied in the field of measuring 5′-nucleotidase activity and 5′-nucleotidase diagnostic kit, can solve the problem of poor accuracy, complex methods, and difficulty in popularization and application and other problems to achieve the effect of good accuracy and accurate test results

Inactive Publication Date: 2006-04-19
王尔中
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  • 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

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0101] Embodiment one (single agent)

[0102] The 5'-nucleotidase diagnostic kit of the present embodiment comprises:

[0103] Buffer 80mmol / l

[0104] Inosine monophosphate 1mmol / l

[0105] Monovalent phosphate 2mmol / l

[0106] Nucleoside phosphatase 5000U / l

[0107] Xanthine oxidase 5000U / l

[0108] Peroxidase 5000U / l

[0109] Stabilizer 50% (total volume)

[0110] 4-amino antiarsenic 2mmol / l

[0111] Carbolic acid 10mmol / l

[0112] Set on the automatic biochemical analyzer: temperature 37, reaction time 10 minutes, test main wavelength 495-505nm, test secondary 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.

[0113] After the sample and reagents are added and allowed to mix, the following reactions occur:

[0114] Inosine monophosphate + water 5 nucleotidase Inosine + Phosphate

[0115] Inos...

Embodiment 2

[0121] Embodiment two (two doses)

[0122] The 5'-nucleotidase diagnostic reagent of the present embodiment has:

[0123] Reagent I

[0124] Buffer 100mmol / l

[0125] Monovalent phosphate 5mmol / l

[0126] Nucleoside phosphatase 8000U / l

[0127] Xanthine oxidase 8000U / l

[0128] Peroxidase 8000U / l

[0129] Stabilizer 50% (total volume)

[0130] 4-amino antiarsenic 1mmol / l

[0131] Reagent II

[0132] Buffer 100mmol / l

[0133] Inosine monophosphate 3mmol / l

[0134] Stabilizer 30mmol / l

[0135] 2,4,6-tribromo-3-hydroxy-benzenesulfonic acid 1mmol / l

[0136] 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 600nm, 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.

[0137] The spec...

Embodiment 3

[0145] Embodiment three (three doses)

[0146] The 5′-nucleotidase diagnostic reagent of the present embodiment is three doses, including:

[0147] Reagent I

[0148] Buffer 120mmol / l

[0149] Monovalent phosphate 8mmol / l

[0150] Stabilizer 50mmol / l

[0151] 3-methyl-2-benzothiazolone hydrazone 2mmol / l

[0152] Reagent II

[0153] Buffer 120mmol / l

[0154] Nucleoside phosphatase 12000U / l

[0155] Xanthine oxidase 12000U / l

[0156] Peroxidase 12000U / l

[0157] Stabilizer 50% (total volume)

[0158] Reagent III

[0159] Buffer 120mmol / l

[0160] Inosine monophosphate 5mmol / l

[0161] Stabilizer 20mmol / l

[0162] Dimethylaniline 2mmol / l

[0163] 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...

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Abstract

A reagent kit for diagnosing 5í»-nucleotidase is composed of buffer liquid, inosine monophosphate, monovalence phosphate, nucleotide phosphatase, xanthine oxidase, peroxidase, stabilizer and reduced chromogen group. A process for measuring the activity of 5í»-nucleotidase includes such steps as proportionally mixing specimen with reagents, enzyme coupling reaction and biochemically analyzing the final resultant to determine the variation in light absorptivity of master wavelength and in turn the activity of 5í»-nucleotidase. Its advantages are high sensitivity and precision, and no pollution.

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

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/42
Inventor 王尔中
Owner 王尔中
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