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Monoamine oxidase diagnostic reagent (kit) and method for determining activity concentration of monoamine oxidase

A monoamine oxidase and reagent 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: 2010-08-04
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] Reduced coenzyme 0.25mmol / L

[0045] Nitrite reductase (EC 1.7.2.2) 10000U / L

[0046] Nitrite reductase (EC 1.7.1.4) 10000U / L

[0047] Amine compound 3mmol / L

[0048] Ferricytochrome C 5mmol / L

[0049] Calcium chloride 2mmol / 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 m...

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] Reduced coenzyme 0.25mmol / L

[0059] Amine compound 3mmol / L

[0060] Ferricytochrome C 5mmol / L

[0061] Calcium chloride 2mmol / L

[0062] Reagent 2

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

[0064] Stabilizer 500mmol / L

[0065] Nitrite reductase (EC 1.7.2.2) 10000U / L

[0066] Nitrite reductase (EC 1.7.1.4) 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 s...

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] Reduced coenzyme 0.25mmol / L

[0076] Amine compound 3mmol / L

[0077] Ferricytochrome C 5mmol / L

[0078] Calcium chloride 2mmol / L

[0079] Reagent 2

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

[0081] Stabilizer 500mmol / L

[0082] Nitrite reductase (EC 1.7.1.4) 10000U / L

[0083] Reagent 3

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

[0085] Stabilizer 500mmol / L

[0086] Nitrite reductase (EC 1.7.2.2) 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 biochemic...

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Abstract

The invention relates to a monoamine oxidase diagnosing / determining reagent (kit) utilizing the technologies of the indirect enzymatic recycling amplification method, the enzymatic-colorimetric method and the enzyme-link method and simultaneously also relates to a method, constitution and components of a reagent for determining the activity concentration of the monoamine oxidase, belonging to the technical field of medical testing and determination. The reagent (kit) comprises the following principle components: a buffer solution, a reduced coenzyme, an amine compound, ferricytochrome C, calcium chloride, a nitrite reductase (EC 1.7.2.2), another nitrite reductase (EC 1.7.1.4) and a stabilizer; and samples and the reagent are mixed according to a certain volume ratio so as to generate a series of enzymatic reactions, and reactants are placed under an ultraviolet / visible light analyzer so as to detect the ascending velocity of absorbency of a part with dominant wavelength being 340 nm, thereby obtaining the activity concentration of the monoamine oxidase through measurement and calculation.

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): G01N21/33G01N35/00C12Q1/26
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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