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Determination of 5'-nucleotidase activity and its diagnostic reagent kit of 5'-nucleotidase

A diagnostic kit and nucleotidase technology are applied in the field of measuring 5′-nucleotidase activity and 5′-nucleotidase diagnostic kits, and can solve the problem of being unsuitable for popularization and application, environment polluted by strong acid, and difficult to popularize. Application and other issues, to achieve the effect of low test cost, accurate test results, and easy promotion and application

Inactive Publication Date: 2006-05-31
SUZHOU ANJ BIOTECHNOLOGY CO LTD
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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

[0063] Embodiment one (single agent)

[0064] The 5'-nucleotidase diagnostic kit of the present embodiment comprises:

[0065] Buffer 80mmol / l

[0066] Uridine monophosphate 2mmol / l

[0067] Reduced coenzyme 0.2mmo / l

[0068] Uridine phosphorylase 5000U / l

[0069] Dihydrothymidine dehydrogenase 5000U / l

[0070] Stabilizer 50% (total volume)

[0071] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, test main wavelength 340nm, test sub-wavelength above 405nm, the volume ratio of the tested 5'-nucleotidase sample to the reagent is 1 / 25, The reaction direction is negative reaction, the delay time is 1 minute, the detection time is 2 minutes, and the theoretical K value is -4180.

[0072] After the sample and reagents are added and allowed to mix, the following reactions occur:

[0073] Uridine monophosphate + water 5 nucleotidase Uridine + Phosphate

[0074] Uridine + Phosphate uridine phosphorylase Uracil + ribose phospha...

Embodiment 2

[0079] Embodiment two (two doses)

[0080] The 5'-nucleotidase diagnostic reagent of the present embodiment has:

[0081] Reagent I

[0082] Buffer 100mmol / l

[0083] Reduced coenzyme 0.25mmo / l

[0084] Uridine phosphorylase 8000U / l

[0085] Dihydrothymidine dehydrogenase 8000U / l

[0086] Stabilizer 50% (total volume)

[0087] Reagent II

[0088] Buffer 100mmol / l

[0089] Uridine monophosphate 6mmol / l

[0090] Stabilizer 20mmol / l

[0091] 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 340nm, the secondary wavelength of the test is above 405nm, and the volume ratio of the tested 5′-nucleotidase sample to the reagent is 1 / 25, the reaction direction is negative reaction, the delay time is 1 minute, the detection time is 2 minutes, and the theoretical K value is -4180.

[0092] The specific measurement steps are:

[0093] Uridine monophosphate + water 5 nucleotidas...

Embodiment 3

[0099] Embodiment three (three doses)

[0100] The 5′-nucleotidase diagnostic reagent of the present embodiment is three doses, including:

[0101] Reagent I

[0102] Buffer 120mmol / l

[0103] Reduced coenzyme 0.3mmo / l

[0104] Stabilizer 20mmol / l

[0105] Reagent II

[0106] Buffer 120mmol / l

[0107] Uridine phosphorylase 10000U / l

[0108] Dihydrothymidine dehydrogenase 10000U / l

[0109] Stabilizer 50% (total volume)

[0110] Reagent III

[0111] Buffer 120mmol / l

[0112] Uridine monophosphate 10mmol / l

[0113] Stabilizer 20mmol / l

[0114] Set on the automatic biochemical analyzer: temperature 25°C, reaction time 20 minutes, test main wavelength 340nm, test sub-wavelength above 405nm, the volume ratio of the tested 5'-nucleotidase sample to the reagent is 1 / 25, The reaction direction is negative reaction, the delay time is 1 minute, the detection time is 2 minutes, and the theoretical K value is -4180.

[0115] The specific measurement steps are:

[0116] Ur...

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PUM

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Abstract

The invention is about a method of measuring the activation of 5í»-phosphonuclease, and it also concerns the reagent box of 5í»-phosphonucleasediagnosis. This invention belongs to the field of medical testing and measuring technology. The reagent box is consisted of buffer solution, uridine monophosphate, reduced coenzyme, uridine phosphorylase, dihydrothymine dehyddrogenase and stabilizer. Firstly, we cause an enzyme-coupled reaction through mixing the sample and the reagent according to a certain proportion of volume; secondly, put the final reactant under the biochemical analyzer and test the absorbance variational situation (speed) of dominant wavelength; then we can get theactivation of 5í»-phosphonuclease. By using this invention, we can get the necessary measuring result with high sensitiveness and fine precision through biochemical analyzer, and the result would not be contaminated by material of internal and exogenous sources. Thus, this method can be conveniently promoted and applied.

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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IPC IPC(8): C12Q1/32
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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