Adenosine triphosphate (ATP ) fluorescence detection method

Abstract

The invention relates to the technical field of product detection, and provides an ATP fluorescence detection method with strong fluorescence intensity, sensitive detection, high fluorescence reaction rate per unit time, substrate recycling, and increased fluorescence signal output cycle, including the following processing steps: (1) Sample collection; (2) Add firefly luciferase and firefly luciferin regenerating enzyme luciferin regenerating enzyme complex structure, reaction substrate firefly luciferin, magnesium ions, cysteine ​​into the reaction vessel, pass in an appropriate amount of solvent, fully Mix and dissolve to obtain the reaction system mixture solution; (3) The substrate mixture solution fully dissolves the ATP in the sample to be tested, and under the condition of atmospheric oxygen, the reaction is promoted to produce oxidized fluorescein and light; (4) Oxidized fluorescence Under the action of firefly luciferin regenerating enzyme luciferin regenerating enzyme and the substrate cysteine, the luciferin is reduced to firefly luciferin, which promotes the reaction to continue; (5) collecting light signals: the light generated by the reaction in step (3) is carried out collection.

Description

technical field [0001] The invention relates to the technical field of product detection, in particular to an ATP fluorescence detection method. Background technique [0002] Adenosine triphosphate (ATP) exists in the cells of organisms from microorganisms to higher animals and plants. Its main function is to provide energy and participate in the metabolism of fat, protein, sugar and nucleic acid in the body. It is an important source of energy for the body. Functionally, it has an irreplaceable role. The rapid and efficient determination of ATP is of great significance for the study of the physiological activity and metabolic process of cells and even the body, drug sensitivity experiments and food hygiene monitoring. [0003] The rapid detection of food safety is a real-time sampling of food / beverage, and the ATP content of the sample can reflect the total number of microorganisms in the sample, and then indicate the freshness of the food / beverage. The ATP detection prod...

AI Technical Summary

Problems solved by technology

Reactions such as figure 1 As shown, this ATP detection product has limitations, which are mainly affected by factors such as the stability of firefly fluorescent protein, the half-life of fluorescein, and too weak fluorescence intensity, resulting in insufficient sensitivity, and the effective working period of the detection reagent is too short

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