Efficient plasma, erythrocyte and lymphocyte NAD<+> detection method

Abstract

The invention provides an NAD<+> detection method, which can rapidly and sensitively detect the NAD<+> level in plasma, erythrocyte and lymphocyte, and belongs to the field of molecular biology and medical detection. According to the method, NAD<+> is reduced into NADH under the action of alcohol dehydrogenase (ADH) by adopting an enzymatic cycling method principle, MTT is reduced into blue formazan under the action of an electronic coupling reagent PMS by the NADH, the formazan generation rate is detected by an enzymatic kinetics method, and the amount of the NAD<+> in a sample can be obtained by combining an NAD<+> standard curve. The method mainly comprises the following steps: sample preparation, reagent preparation, r-SBFA preparation, beta-NAD standard substance preparation, reaction liquid preparation, NAD<+> extraction, interfering substance removal, NAD<+> detection and data calibration. The method is suitable for detecting the NAD<+> level in mammalian plasma, erythrocyte and lymphocyte, the unique sample treatment method solves the problem that NAD<+> is difficult to detect due to multiple interfering substances in a sample, and the method has the advantages of low detection limit, short time and low cost, and is suitable for high-throughput detection, easy to apply and popularize and high in practicability.

Description

technical field [0001] The invention relates to the fields of molecular biotechnology and medical detection, in particular to a highly efficient blood NAD + A detection method that can comprehensively evaluate NAD in mammalian plasma, red blood cells, and lymphocytes + Level. Background technique [0002] Nicotinamide adenine dinucleotide (NAD + ) is an important member of the pyridine nucleotide family. It is not only a classic coenzyme that mediates a variety of redox reactions, but also a co-substrate for enzymes such as Sirtuins, PARPs, and CD38. play a key role in stability. Studies have shown that NAD in the body + Levels decline with age, and this decline plays a crucial role in the development of aging and age-related diseases. CurrentNAD + As an important target for delaying aging and prolonging lifespan, it plays an increasingly prominent role in biopharmacology and future disease research, so the development of a reliable NAD + The detection method is criti...

AI Technical Summary

Problems solved by technology

[0004] The present invention aims to solve the problem of rapid and accurate determination of NAD in mammalian plasma, red blood cells and lymphocytes + Level, the unique sample processing method solves the interfering substance NAD in the sample + Problems that are difficult to detect, using the present invention to detect NAD + Level, low detection limit, short time-consuming, low cost, suitable for high-throughput detection

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