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Determination of 5'-nucleotidase activity and its 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 the activity of 5′-nucleotidase, can solve problems that are not suitable for promotion and application, strong acid pollutes the environment, and is difficult to promote. 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

[0062] Example one (single dose)

[0063] The 5'-nucleotidase diagnostic kit of this embodiment includes:

[0064] Buffer 80mmol / l

[0065] Inosine monophosphate 2mmol / l

[0066] Oxidized coenzyme 2mmo / l

[0067] Inosine nucleosidase 5000U / l

[0068] Ribose 1-dehydrogenase 5000U / l

[0069] Stabilizer 50% (total volume)

[0070] Set on the automatic biochemical analyzer: temperature 37℃, reaction time 10 minutes, test main wavelength 340nm, test sub-wavelength above 405nm, the volume ratio of the tested 5′-nucleotidase sample to reagent is 1 / 25, The reaction direction is positive reaction, the delay time is 1 minute, the detection time is 2 minutes, and the theoretical K value is -4180.

[0071] After adding the sample and reagents and mixing them, the following reactions occur:

[0072] Inosine Monophosphate + Water 5′-nucleotidase Inosine + Monophosphate

[0073] Inosine + water Inosine nucleosidase Ribose + Uracil

[0074] Ribose + Oxidized Coenzyme + Water Ribose 1-d...

Embodiment 2

[0077] Example two (double dose)

[0078] The 5′-nucleotidase diagnostic reagents of this embodiment include:

[0079] Reagent I

[0080] Buffer 100mmol / l

[0081] Oxidized coenzyme 4mmo / l

[0082] Inosine nucleosidase 8000U / l

[0083] Ribose 1-dehydrogenase 8000U / l

[0084] Stabilizer 50% (total volume)

[0085] Reagent II

[0086] Buffer 100mmol / l

[0087] Inosine monophosphate 6mmol / l

[0088] Stabilizer 20mmol / l

[0089] When measuring the activity of 5'-nucleotidase, the temperature is controlled at 30℃, the reaction time is 15 minutes, the main wavelength of the test is 340nm, and the subwavelength is above 405nm. 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, the detection time is 2 minutes, and the theoretical K value is -4180.

[0090] The specific measurement steps are:

[0091] Inosine Monophosphate + Water 5′-nucleotidase Inosine + Monophosphate

[0092] Inos...

Embodiment 3

[0096] Example three (three doses)

[0097] The 5′-nucleotidase diagnostic reagent in this embodiment consists of three doses, including:

[0098] Reagent I

[0099] Buffer 120mmol / l

[0100] Oxidized coenzyme 6mmo / l

[0101] Stabilizer 20mmol / l

[0102] Reagent II

[0103] Buffer 120mmol / l

[0104] Inosine nucleosidase 10000U / l

[0105] Ribose 1-dehydrogenase 10000U / l

[0106] Stabilizer 50% (total volume)

[0107] Reagent III

[0108] Buffer 120mmol / l

[0109] Inosine monophosphate 10mmol / l

[0110] Stabilizer 20mmol / l

[0111] Set on the automatic biochemical analyzer: temperature 25℃, reaction time 20 minutes, test main wavelength 340nm, test sub-wavelength above 405nm, the volume ratio of the tested 5′-nucleotidase sample to reagent is 1 / 25, The reaction direction is positive reaction, the delay time is 1 minute, the detection time is 2 minutes, and the theoretical K value is -4180.

[0112] The specific measurement steps are:

[0113] Inosine Monophosphate + Water 5′-nucl...

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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, carnine monophosphate, oxidized coenzyme, carnine nucleotide, ribose1-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 the activity of 5'-nucleotidase, and at the same time, the invention also relates to a 5'-nucleotidase diagnostic kit for realizing the method, which belongs to the technical field of medical testing and determination. Background technique [0002] Medical research shows that the increase of 5'-nucleotidase (5NT) is mainly seen in obstructive yellow bile, but also in liver cancer and hepatitis. In cholestasis complicated by cholangitis, primary and secondary biliary cirrhosis and chronic hepatitis, the increase rate of 5NT is higher than that of alkaline phosphatase; the sensitivity of increase of 5NT is higher than that of alkali in liver tumors and liver granuloma性phosphatase. Because the 5'-nucleotidase activity has no physiological increase, it is not only more sensitive than alkaline phosphatase, but also specific for diagnosing infant liver disease and hepatic cholestasis of pregnancy. Therefore, measuri...

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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