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Monoamine oxidase diagnostic reagent (kit) and monoamine oxidase active concentration measuring method

A monoamine oxidase and pyruvate oxidase technology is applied in the field of medical testing and determination, and can solve the problems of inability to use automatic biochemical analyzer analysis, limited practicability, difficult determination and the like

Inactive Publication Date: 2010-08-04
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

[0041] The monoamine oxidase diagnosis / measurement reagent of this embodiment is a single reagent, including:

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

[0043] Stabilizer 500mmol / L

[0044] Coenzyme 3mmol / L

[0045] Pyruvate oxidase 12000U / L

[0046] Coenzyme A-disulfide reductase 10000U / L

[0047] Benzylamine 5mmol / L

[0048] Sodium bicarbonate 12mmol / L

[0049] Acetyl-CoA 3mmol / 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 ≤ 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 ...

Embodiment 2

[0054] The monoamine oxidase diagnosis / measurement 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] Coenzyme 3mmol / L

[0059] Benzylamine 5mmol / L

[0060] Sodium bicarbonate 12mmol / L

[0061] Acetyl-CoA 3mmol / L

[0062] Reagent 2

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

[0064] Stabilizer 500mmol / L

[0065] Pyruvate oxidase 12000U / L

[0066] Coenzyme A-disulfide reductase 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 ≤ 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 i...

Embodiment 3

[0071] The monoamine oxidase diagnosis / measurement reagent of the present embodiment is three reagents, comprising:

[0072] Reagent 1

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

[0074] Stabilizer 50mmol / L

[0075] Coenzyme 3mmol / L

[0076] Benzylamine 5mmol / L

[0077] Sodium bicarbonate 12mmol / L

[0078] Acetyl-CoA 3mmol / L

[0079] Reagent 2

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

[0081] Stabilizer 500mmol / L

[0082] Coenzyme A-disulfide reductase 10000U / L

[0083] Reagent 3

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

[0085] Stabilizer 500mmol / L

[0086] Pyruvate oxidase 12000U / 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: temperature 37°C, r...

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Abstract

The invention relates to a monoamine oxidase diagnostic / measuring reagent (kit) which utilizes enzyme colorimetry and enzyme-linked method technologies, and meanwhile, the invention also relates to a method for measuring monoamine oxidase active concentration and compositions and components of the reagent, which belong to the technical field of medical inspection and measurement. The main components of the reagent (kit) of the invention comprise a buffer solution, coenzyme, an amine compound, sodium bicarbonate, acetyl coenzyme A, pyruvate oxidase, coenzyme A-disulfide reductase and a stabilizer; and the method comprises the following steps of: enabling a sample and the reagent to generate a series of enzymatic reactions through mixing the sample and the reagent according to a certain volume ratio, then placing the reactants under an ultraviolet / visible light analyzer and detecting the rising speed of the absorbance at a position with the dominant wavelength of 340nm to measure and calculate the active concentration magnitude of 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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IPC IPC(8): G01N21/33G01N35/00C12Q1/26C12Q1/00
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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