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5'nucleotidase diagnostic kit and method for measuring activity concentration of 5'nucleotidase

A nucleotidase and nucleoside monophosphate technology, which is applied in the field of medical/food/environmental testing and determination, can solve the problems of strong acid pollution, difficulty in popularization and application, and poor accuracy.

Inactive Publication Date: 2009-12-23
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

[0043] The 5' nucleotidase diagnostic reagent of the present embodiment is a single reagent, comprising:

[0044] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0045] Stabilizer 500mmol / L

[0046] Coenzyme 3mmol / L

[0047] Glutamine Synthetase 6000U / L

[0048] Glutamate synthase 8000U / L

[0049] Nucleoside monophosphate 9mmol / L

[0050] Glutamine 12mmol / L

[0051] Adenosine diphosphate 6mmol / L

[0052] After the reagents are all dissolved and prepared, they are divided into bottles and freeze-dried to make dry powder reagents; before use, add purified water and reconstitute for use.

[0053] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, initial absorbance ≤ 0.1, test main wavelength 340nm, test secondary wavelength 405nm, the volume ratio of the tested 5' nucleotidase sample to the reagent is 1 / 25, the reaction direction is positive reaction (rising reaction), the delay time is about 1 minute, the detection...

Embodiment 2

[0056] The 5' nucleotidase diagnostic reagent of the present embodiment is a double reagent, comprising:

[0057] Reagent 1

[0058] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0059] Stabilizer 50mmol / L

[0060] Coenzyme 3mmol / L

[0061] Nucleoside monophosphate 9mmol / L

[0062]Glutamine 12mmol / L

[0063] Adenosine diphosphate 6mmol / L

[0064] Reagent 2

[0065] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0066] Stabilizer 500mmol / L

[0067] Glutamine Synthetase 6000U / L

[0068] Glutamate synthase 8000U / L

[0069] After the reagents are all dissolved and prepared, they are divided into bottles to make liquid double reagents, which can be used directly.

[0070] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, initial absorbance ≤ 0.1, test main wavelength 340nm, test secondary wavelength 405nm, tested 5' nucleotidase sample and reagent 1, reagent 2 The volume ratio is 2 / 20 / 5, th...

Embodiment 3

[0073] The 5' nucleotidase diagnostic reagent of the present embodiment is three reagents, comprising:

[0074] Reagent 1

[0075] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0076] Stabilizer 50mmol / L

[0077] Coenzyme 3mmol / L

[0078] Nucleoside monophosphate 9mmol / L

[0079] Glutamine 12mmol / L

[0080] Adenosine diphosphate 6mmol / L

[0081] Reagent 2

[0082] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0083] Stabilizer 500mmol / L

[0084] Glutamate synthase 8000U / L

[0085] Reagent 3

[0086] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0087] Stabilizer 500mmol / L

[0088] Glutamine Synthetase 6000U / L

[0089] After all the reagents are dissolved and prepared, they are divided into bottles to make liquid three reagents, which can be used directly.

[0090] When measuring the concentration of 5' nucleotidase activity, set it on the automatic biochemical analyzer: temperature 37°C, reaction...

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PUM

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Abstract

The invention relates to a 5'nucleotidase diagnostic kit utilizing technologies of an enzymatic-colorimetric method and an enzyme-link method, also relates to a method and a principle of measuring activity concentration of 5'nucleotidase and compositions and components of reagents, and belongs to the technical field of testing and measuring of medical science. The kit mainly comprises the following compositions: buffer solution, coenzyme, nucleoside monophosphate, glutamine, adenosine diphosphate, glutamine synthetases, glutamate synthetase and stabilizer; samples are mixed with the reagents in certain volume ratio to perform a series of enzymatic reactions; then reactants are placed under a UV / visible analyzer; and the descending speed of absorbance is tested at the position where dominant wave length is 340nm so as to measure and calculate the activity concentration of the 5'nucleotidase.

Description

technical field [0001] The invention relates to a 5' nucleotidase diagnostic kit, and at the same time, the invention also relates to a method for measuring the activity concentration of 5' nucleotidase, which belongs to the technical field of medical / food / environmental testing 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 ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31G01N21/25C12Q1/68C12Q1/00
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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