Monoamine oxidase diagnostic reagent (kit) and activity concentration measurement method of same

A monoamine oxidase and glycine oxidase technology, which is applied in the field of medical testing and determination, can solve the problems of limited practicability, difficulty in determination, and inability to use automatic biochemical analyzer analysis.

Inactive Publication Date: 2010-06-23
SUZHOU ANJ BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The principle of the benzylamine method is that benzylamine generates benzylaldehyde under the action of monoamine oxidase, and then reacts with dinitrophenylhydrazine under the condition of strong alkaline sodium hydroxide to generate aldehyde phenylhydrazone, which is difficult to measure on a biochemical instrument; The p-benzylamine-β-azonaphthol method needs to be extracted with cyclohexane, which limits the practicability of the method and cannot be analyzed by a fully automatic biochemical analyzer

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] The monoamine oxidase diagnosis / measurement reagent of this embodiment is a single reagent, including:

[0040] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0041] Stabilizer 500mmol / L

[0042] Coenzyme 3mmol / L

[0043] Glycine oxidase 6000U / L

[0044] Alanine dehydrogenase 8000U / L

[0045] Amine compound 4mmol / L

[0046] Pyruvate 12mmol / L

[0047] Hydrogen peroxide 12mmol / L

[0048] 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.

[0049] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, initial absorbance ≤ 0.1, test main wavelength 340nm, test subwavelength 405nm, the volume ratio of the tested monoamine oxidase sample to the reagent is 1 / 25, the reaction The direction is positive reaction (rising reaction), the delay time is about 1 minute, the detection time is abo...

Embodiment 2

[0052] The monoamine oxidase diagnosis / measurement reagent of this embodiment is a double reagent, comprising:

[0053] Reagent 1

[0054] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0055]Stabilizer 50mmol / L

[0056] Coenzyme 3mmol / L

[0057] Amine compound 4mmol / L

[0058] Pyruvate 12mmol / L

[0059] Hydrogen peroxide 12mmol / L

[0060] Reagent 2

[0061] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0062] Stabilizer 500mmol / L

[0063] Glycine oxidase 6000U / L

[0064] Alanine dehydrogenase 8000U / L

[0065] After the reagents are all dissolved and prepared, they are divided into bottles to make liquid double reagents, which can be used directly.

[0066] 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 monoamine oxidase sample to reagent 1 and reagent 2 is 2 / 20 / 5, ...

Embodiment 3

[0069] The monoamine oxidase diagnosis / measurement reagent of the present embodiment is three reagents, comprising:

[0070] Reagent 1

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

[0072] Stabilizer 50mmol / L

[0073] Coenzyme 3mmol / L

[0074] Amine compound 4mmol / L

[0075] Pyruvate 12mmol / L

[0076] Hydrogen peroxide 12mmol / L

[0077] Reagent 2

[0078] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0079] Stabilizer 500mmol / L

[0080] Alanine dehydrogenase 8000U / L

[0081] Reagent 3

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

[0083] Stabilizer 500mmol / L

[0084] Glycine oxidase 6000U / L

[0085] After all the reagents are dissolved and prepared, they are divided into bottles to make liquid three reagents, which can be used directly.

[0086] When measuring the activity concentration of monoamine oxidase, set it on the automatic biochemical analyzer: temperature 37°C, reaction ...

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Abstract

The invention relates to a monoamine oxidase diagnosis / measurement reagent (kit) utilizing an indirect enzymatic recycling amplification method, an enzyme-colorimetric method and an enzyme coupling method, a method for measuring the activity concentration of the monoamine oxidase, a composition and components of the reagent, belonging to the technical field of medical inspection and measurement. The reagent (kit) comprises the following main components: a buffer solution, a coenzyme, an amine compound, pyruvic acid, hydrogen peroxide, a glycine oxidase, an alanine dehydrogenase and a stabilizing agent. The method comprises the following steps of: mixing a sample with the reagent according to a certain volume ratio so as to ensure that the sample and the reagent have a series of enzymatic reactions; placing an obtained reactant under an ultraviolet / visible light analyzer; and detecting the ascending speed of absorbance at a 340nm part of a main wavelength so as to measure the activity concentration of the monoamine oxidase.

Description

technical field [0001] The invention relates to a monoamine oxidase diagnosis / measurement reagent (box), and at the same time, the invention also relates to a method for measuring the activity concentration of the monoamine oxidase, which belongs to the technical field of medical examination and measurement. Background technique [0002] In the prior art, the methods for measuring the activity of monoamine oxidase (MAO) mainly include: fluorescence method, immunosuppression method and chemical spectrophotometry. The fluorescence method uses β-phenylethylamine as a substrate to detect monoamine oxidase, and the basis is that the reaction rate of β-phenylethylamine is the highest at 10-100 mg, which is higher than that of benzylamine and 5-hydroxytryptamine. The immunological method uses monoamine oxidase antibody to react with monoamine oxidase to measure the monoamine oxidase isoenzyme formed after the reaction. Currently, the most widely used monoclonal antibodies are MAO-A...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31G01N21/17G01N33/48C12Q1/26
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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