Monoamine oxidase diagnosing reagent kit and method for measuring monoamine oxidase activity concentration

A technology of monoamine oxidase and diagnostic kits, which is applied in the field of medical inspection and measurement, and can solve the problems of inability to apply automatic biochemical instruments, limited practicability, and measurement difficulties.

Inactive Publication Date: 2008-01-02
SUZHOU ANJ BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, benzylamine and p-benzylamine-β-azonaphthol are mostly used as substrates for the determination of monoamine oxidase, but the p-benzylamine-β-azonaphthol benzylamine method needs to be extracted with cycloethane, which limits the practicality of the method The principle of the benzylamine method is that benzylamine generates benzaldehyde under the action of monoamine oxidase, and then reacts with dinitrophenylhydrazine under the condition of strong alkaline sodium hydroxide to generate aldehyde benzene Hydrazone is also difficult to measure on a biochemical analyzer

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] The monoamine oxidase diagnostic reagent of this embodiment is a single reagent, comprising:

[0045] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0046] Stabilizer 50mmol / L

[0047] Reduced coenzyme 0.25mmol / L

[0048] Glutamate dehydrogenase 50000U / L

[0049] Glutamate oxidase 10000U / L

[0050] α-ketoglutarate 16mmol / L

[0051] 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.

[0052] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, initial absorbance 1.8±0.7, test main wavelength 340nm, test secondary wavelength 405nm, the volume ratio of the sample to be tested is 1 / 25, the reaction The direction is negative reaction (falling reaction), the detection method is the rate method, the delay time is about 1 minute, and the detection time is about 2 minutes.

[0053] After...

Embodiment 2

[0055] The monoamine oxidase diagnostic reagent of this embodiment is a double reagent, comprising:

[0056] Reagent 1

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

[0058] Stabilizer 50mmol / L

[0059] Reduced coenzyme 0.25mmol / L

[0060] α-ketoglutarate 12mmol / L

[0061] Reagent 2

[0062] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0063] Stabilizer 50mmol / L

[0064] Glutamate dehydrogenase 40000U / L

[0065] Glutamate oxidase 15000U / L

[0066] After the reagents are all dissolved and prepared, they are divided into bottles to make liquid double reagents, which can be used directly.

[0067] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, initial absorbance 1.8±0.7, test main wavelength 340nm, test secondary wavelength 405nm, the volume ratio of the tested sample to reagent 1 and reagent 2 is 2 / 20 / 5, the reaction direction is negative reaction (decreasing reaction), the dete...

Embodiment 3

[0070] The monoamine oxidase diagnostic reagent of this embodiment is three reagents, comprising:

[0071] Reagent 1

[0072] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0073] Stabilizer 50mmol / L

[0074] Reduced coenzyme 0.25mmol / L

[0075] α-ketoglutarate 10mmol / L

[0076] Reagent 2

[0077] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0078] Stabilizer 50mmol / L

[0079] Glutamate dehydrogenase 60000U / L

[0080] Reagent 3

[0081] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0082] Stabilizer 50mmol / L

[0083] Glutamate oxidase 20000U / L

[0084] After all the reagents are dissolved and prepared, they are divided into bottles to make liquid three reagents, which can be used directly.

[0085] When measuring the activity concentration of monoamine oxidase, set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, initial absorbance 1.8±0.7, test main wavelength 340...

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PUM

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Abstract

The invention relates to a monoamine oxidase diagnosis reagent box of Enzymatic Recycling Method, and the invention also relates to method principle for detecting the solution of monoamine oxidase, constitute and component of reagent, which belongs to the technical field of medical checking measurements. The reagent box in the invention can be dry powder state, and used after dissolution; it also can be formulated to be liquid agent for direct usage. The component of reagent box mainly contains: buffering liquid, alpha- ketoglutaric acid, glutamate dehydrogenase EC 1.4.1.2, EC 1.4.1.3, EC 1.4.1.4, Glutamate oxidase EC 1.4.3.7, EC 1.4.3.11, reduction type coenzyme and stabilizer, and the component can be matched to form single-reagent reagent box, two-reagent reagent box, and three- reagent box; The enzymatic reaction is generated between sample and reagent by mixing sample and reagent with definite volume, then the reactants is placed under the ultraviolet/visible light analyzer to detect ascend degree of absorbance at main wavelength of 340nm, so concentration of monoamine oxidase can be calculated. The invention can obtain the detecting result by ultraviolet/visible light analyzer, and the sensibility is high, precision is well, and easy to be spread.

Description

technical field [0001] The invention relates to a diagnostic kit for monoamine oxidase, and at the same time, the invention also relates to a method for measuring the activity concentration of monoamine oxidase, which belongs to the technical field of medical examination and measurement. Background technique [0002] The determination of monoamine oxidase (MAO) can be divided into chemical spectrophotometry, fluorescence method, immunosuppressive method and enzymatic method which is now applying for a patent. Generally, chemical spectrophotometry is used. The substrates of monoamine oxidase are benzylamine and p-benzylamine-β-azonaphthol. Monoamine oxidase can also be used to catalyze the release of H from amines. 2 o 2 Oxidative chromogens such as 10-N-methylcarbamoyl-3,7-dimethylamino-10-hydro-phenothiazine (MCDP). In addition, the reaction substrates of monoamine oxidase include butylamine, amylamine, b-phenylethylamine, tyramine (3-p-hydroxyphenylethylamine 3 -parahdr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31G01N21/78G01N33/50G01N33/52C12Q1/26
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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