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5'nucleotidase diagnosis kit and method for determining 5'nucleotidase activity concentration

A technology of nucleotidase and inosine monophosphate dehydrogenase, which is applied in the field of medical inspection and measurement, and can solve problems such as poor accuracy, isotope pollution, and complicated methods

Inactive Publication Date: 2010-01-06
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

[0037] The 5' nucleotidase diagnostic reagent of the present embodiment is a single reagent, comprising:

[0038] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0039] Stabilizer 500mmol / L

[0040] Coenzyme 3mmol / L

[0041] Inosine monophosphate dehydrogenase 9000U / L

[0042] Phosphate 12mmol / L

[0043] Inosine 8mmol / L

[0044] After all the reagents are dissolved and prepared, they are divided into bottles and freeze-dried to make dry powder reagents;

[0045] Before use, add purified water and reconstitute before use.

[0046] 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 time is about 2 minutes, and the theoretical K value is 4180.

...

Embodiment 2

[0049] The 5' nucleotidase diagnostic reagent of the present embodiment is a double reagent, comprising:

[0050] Reagent 1

[0051] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0052] Stabilizer 50mmol / L

[0053] Coenzyme 3mmol / L

[0054] Phosphate 12mmol / L

[0055] Inosine 8mmol / L

[0056] Reagent 2

[0057] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0058] Stabilizer 500mmol / L

[0059] Inosine monophosphate dehydrogenase 9000U / L

[0060] After the reagents are all dissolved and prepared, they are divided into bottles to make liquid double reagents, which can be used directly.

[0061] 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, the reaction direction is positive reaction (rising reaction), the delay time is...

Embodiment 3

[0064] The 5' nucleotidase diagnostic reagent of the present embodiment is three reagents, comprising:

[0065] Reagent 1

[0066] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0067] Stabilizer 50mmol / L

[0068] Coenzyme 3mmol / L

[0069] Phosphate 12mmol / L

[0070] Reagent 2

[0071] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0072] Stabilizer 500mmol / L

[0073] Inosine 8mmol / L

[0074] Reagent 3

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

[0076] Stabilizer 500mmol / L

[0077] Inosine monophosphate dehydrogenase 9000U / L

[0078] After all the reagents are dissolved and prepared, they are divided into bottles to make liquid three reagents, which can be used directly.

[0079] When measuring the concentration of 5' nucleotidase activity, set it on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, initial absorbance ≤ 0.1, test main wavelength 340nm, test se...

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Abstract

The invention relates to a 5'nucleotidase diagnosis kit using the technology of an enzyme colorimetric method and an ELISA method, a method for determining 5'nucleotidase activity concentration, and composition and components of a reagent, and belongs to the technical field of medical test and determination. The kit comprises the following main components: buffer solution, coenzyme, phosphate radical, inosine, inosine monophosphate dehydrogenase, and a stabilizer. The method comprises the following steps: mixing a sample and the reagent according to a certain volume ratio to perform a series of enzymic reaction, and then placing reactants under a UV / visible light analyzer to test the ascending speed of absorbance at the position where the dominant wave length is 340nm to determine the activity concentration of 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 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. ...

Claims

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Application Information

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IPC IPC(8): G01N21/31G01N21/25C12Q1/68C12Q1/32
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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