Application and method adopting single sulfonic acid tetrazolium salt to detect cell activity

Abstract

The invention relates to the technical field of medical biological detection and provides a cell activity detection method. Color changing of a novel single sulfonic acid tetrazolium salt reagent is utilized to monitor the cell activity in real time. A single sulfonic acid tetrazolium salt detecting system is selected for the testing agent, and the good linear dose relation is taken on the NAD(P)H level and the living cell number. According to the testing agent, through the absorbance value of a single sulfonic acid tetrazolium salt reactant at 450 nm, an EC50 curved line is obtained, and evaluation of the cell activity is conducted. The detecting method is based on the cell high sensitiveness. Single sulfonic acid tetrazolium salt is small in cell toxicity and suitable for a living cell foundation test. The single sulfonic acid tetrazolium salt reagent is mild, and detected cells can be further used for the subsequent test after being rinsed. The single sulfonic acid tetrazolium salt reagent is not prone to reacting with reducing substances, and interference is small. The reagent is wide in application range and is suitable for detection after environmental microorganism enrichment, detection of the activity of algae, fungi and the animal and plant cells and detection of the cells and tissue lesion.

Description

technical field [0001] The invention relates to the technical field of medical biological detection, in particular to the application and method of using monosulfonic acid tetrazolium salt for detecting cell viability. Background technique [0002] NAD(P) + / NAD(P)H is an important coenzyme factor that exists in all organisms, including bacteria, actinomycetes, cyanobacteria, yeasts, molds, fungi, general animals and plants, and participates in various life processes as a reaction medium , including energy metabolism, mitochondrial function, calcium homeostasis, oxidative stress, gene regulation, immune function, aging, and cell death. Therefore, a sensitive and low-toxic detection method based on the cell level to measure changes in NAD(P)H or its related enzyme reactions plays a key role in applications such as scientific research, environmental maintenance, and medical diagnosis. [0003] Several different types of tetrazolium salts have been reported to measure cell vi...

AI Technical Summary

Problems solved by technology

MTT (3(4,5-dimethyl-2-thiazolyl)-2 5-phenyl-2-tetrazolium) is a dye that accepts hydrogen ions, which is produced by TTC in the detection of cell viability assays Formazan has very poor water solubility and needs to be dissolved by adding organic reagents (such as DMSO), which is prone to incomplete dissolution and affects the accuracy of the experiment, and it is easy to take away some cells when absorbing the supernatant.

On the other hand, in addition to being reduced by mitochondrial dehydrogenases, MTT can also be reduced by a variety of other substances, such as serum albumin, L-ascorbic acid, β-mercaptoethanol, glutathione s-transferase, etc., and currently The most commonly used reagent on the market, CCK8 (WST-8), has been experimentally confirmed to be reduced by antioxidants such as β-mercaptoethanol and EGCG, resulting in false positives in cell viability tests

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