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Monoamine oxidase diagnosis kit and method for determining monoamine oxidase activity concentration

A monoamine oxidase and reagent technology, applied in the field of medical testing and determination, can solve the problems of inability to use an automatic biochemical instrument, limited practicability, difficulty in determination and the like

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

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

[0041] The monoamine oxidase diagnostic reagent of this embodiment is a single reagent, comprising:

[0042] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0043] Stabilizer 500mmol / L

[0044] Reduced coenzyme 0.25mmol / L

[0045] Pyruvate oxidase 10000U / L

[0046] Lactate dehydrogenase 10000U / L

[0047] Amine compound 10mmol / L

[0048] Sodium bicarbonate (carbon dioxide) 12mmol / L

[0049] Acetyl phosphate 8mmol / L

[0050] 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.

[0051] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, initial absorbance 1.8±0.7, test main wavelength 340nm, test subwavelength 405nm, volume ratio of tested monoamine oxidase sample to reagent is 1 / 25, The reaction direction is negative reaction (decreasing reaction), the delay time is about 1 minute, the d...

Embodiment 2

[0054] The monoamine oxidase diagnostic reagent of this embodiment is a double reagent, comprising:

[0055] Reagent 1

[0056] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0057] Stabilizer 50mmol / L

[0058] Reduced coenzyme 0.25mmol / L

[0059] Amine compound 10mmol / L

[0060] Sodium bicarbonate (carbon dioxide) 12mmol / L

[0061] Acetyl phosphate 8mmol / L

[0062] Reagent 2

[0063] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0064] Stabilizer 500mmol / L

[0065] Pyruvate oxidase 10000U / L

[0066] Lactate dehydrogenase 10000U / L

[0067] After the reagents are all dissolved and prepared, they are divided into bottles to make liquid double reagents, which can be used directly.

[0068] 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 monoamine oxidase sample to reagent...

Embodiment 3

[0071] The monoamine oxidase diagnostic reagent of this embodiment is three reagents, comprising:

[0072] Reagent 1

[0073] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0074] Stabilizer 50mmol / L

[0075] Reduced coenzyme 0.25mmol / L

[0076] Amine compounds 10mmol / L

[0077] Sodium bicarbonate (carbon dioxide) 12mmol / L

[0078] Acetyl phosphate 8mmol / L

[0079] Reagent 2

[0080] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0081] Stabilizer 500mmol / L

[0082] Lactate dehydrogenase 10000U / L

[0083] Reagent 3

[0084] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0085] Stabilizer 500mmol / L

[0086] Pyruvate oxidase 10000U / L

[0087] After all the reagents are dissolved and prepared, they are divided into bottles to make liquid three reagents, which can be used directly.

[0088] When measuring the activity concentration of monoamine oxidase, set on the automatic biochemical analyzer: tempera...

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Abstract

The invention relates to a monoamine oxidase diagnosis kit using the technology of an enzyme colorimetric method and an ELISA method, a method for determining monoamine oxidase 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, reduced coenzyme, amines, sodium bicarbonate (carbon dioxide), acetyl phosphate, pyruvate oxidase, lactate 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 descending speed of absorbance at the position where the dominant wave length is 340nm to determine the activity concentration of the monoamine oxidase.

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): G01N21/31G01N21/25C12Q1/26
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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