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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, can solve the problems of unsuitable promotion and application, strong acid pollution environment, and accuracy It can achieve the effect of low test cost, accurate test result and easy operation

Inactive Publication Date: 2007-02-28
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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  • Method for measuring 5'-nucleotidase activity and diagnostic reagent kit of 5'-nucleotidase
  • Method for measuring 5'-nucleotidase activity and diagnostic reagent kit of 5'-nucleotidase
  • Method for measuring 5'-nucleotidase activity and diagnostic reagent kit of 5'-nucleotidase

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Embodiment one (single agent)

[0067] The 5'-nucleotidase diagnostic kit of the present embodiment comprises:

[0068] Buffer 80mmol / l

[0069] Adenosine monophosphate 5mmol / l

[0070] 2-ketoglutarate 5mmol / l

[0071] Reduced coenzyme 0.15mmo / l

[0072] Adenosine deaminase 10000U / l

[0073] Glutamate dehydrogenase 100000U / l

[0074] Stabilizer 30% (total volume)

[0075] 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.

[0076] After the sample and reagents are added and allowed to mix, the following reactions occur:

[0077]

[0078]

[0079]

[0080] The final reactant is placed under a biochemical analysis instrument, and ...

Embodiment 2

[0082] Embodiment two (two doses)

[0083] The 5'-nucleotidase diagnostic reagent of the present embodiment has:

[0084] Reagent I

[0085] Buffer 100mmol / l

[0086] 2-ketoglutarate 12mmol / l

[0087] Reduced coenzyme 0.2mmol / l

[0088] Adenosine deaminase 15000U / l

[0089] Glutamate dehydrogenase 150000U / l

[0090] Stabilizer 40% (total volume)

[0091] Reagent II

[0092] Buffer 100mmol / l

[0093] Adenosine monophosphate 12mmol / l

[0094] Stabilizer 30mmo l / l

[0095] When measuring 5'-nucleotidase activity, the temperature is controlled at 30°C, the reaction time is 10 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.

[0096] The specific measurement steps are:

[0097]

[0098]...

Embodiment 3

[0102] Embodiment three (three doses)

[0103] The 5′-nucleotidase diagnostic reagent of the present embodiment is three doses, including:

[0104] Reagent I

[0105] Buffer 120mmol / l

[0106] 2-ketoglutarate 20mmol / l

[0107] Reduced coenzyme 0.3mmo / l

[0108] Stabilizer 50mmol / l

[0109] Reagent II

[0110] Buffer 120mmol / l

[0111] Adenosine deaminase 20000U / l

[0112] Glutamate dehydrogenase 200000U / l

[0113] Stabilizer 50% (total volume)

[0114] Reagent III

[0115] Buffer 120mmol / l

[0116] Adenosine monophosphate 20mmol / l

[0117] Stabilizer 50mmol / l

[0118]Set on the automatic biochemical analyzer: temperature 25°C, reaction time 10 minutes, test main wavelength 340nm, test sub-wavelength above 405nm, volume ratio of tested 5'-nucleotidase sample to 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.

[0119] The specific measurement steps are: ...

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Abstract

The present invention relates to 5'-nucleotidase activity detecting method and 5'-nucleotidase diagnosing reagent kit in medical detecting technology. The 5'-nucleotidase diagnosing reagent kit includes buffering liquid, adenosine monophosphate, 2-ketoglutarate, reductive coenzyme, adenosine deaminase, glutamate dehydrogenase, stabilizer, etc. 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 5'-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

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/32
Inventor 王尔中
Owner 王尔中