Homocysteine diagnosis kit and homocysteine concentration measuring method

Abstract

The invention relates to a homocysteine concentration measuring method. The method includes: oxidizing homocysteine to generate alpha-butanone acid, ammonia and hydrogen sulfide; using ammonia to convert NADH into NAD+ under action of glutamate dehydrogenase; quantifying by measuring light absorbance of NADH at a position of 340nm. The homocysteine diagnosis kit comprises a reagent I and a reagent II, the reagent I contains reduced coenzyme I, glutamate dehydrogenase, tri(2-carboxyethyl) phosphine hydrochloride, alpha-ketoglutaric acid, surfactant, buffer solution and stabilizer, and the reagent II contains HCYase, phosphopyridoxal, buffer solution and stabilizer. The homocysteine diagnosis kit and the homocysteine concentration measuring method have the advantages that the kit for detecting homocysteine in human serum through two liquid reagents is built, and an additional fluorescence detector is not needed, so that convenience is brought to detection of plenty of samples; the method is free of interference by pyruvic acid, methionine and cysteine in the samples and lower in cost.

Description

technical field [0001] The invention relates to a homocysteine ​​diagnostic kit and a method for determining the concentration of homocysteine. Background technique [0002] Homocysteine ​​α, γ-lyase (homocysteine ​​α, γ-lyases, HCYase, EC4.4.1.11) is a pyridoxal phosphate (PLP) dependent enzyme, also known as L-methionine γ-lyase, belongs to Gamma protein family. It can catalyze the α, γ-elimination reaction and γ-substitution reaction of L-methionine and homocysteine, and catalyze the α, β elimination reaction and β-substitution reaction of L-cysteine ​​and its derivatives. The enzyme that catalyzes this type of reaction was first discovered in Escherichia coli, named Methionine lyase (Methioninelyase), and now it is named Methionine γ-lyase and found in a variety of organisms, such as Pseudomonas putida, vaginal hair drop Insects, Aeromonas, Arabidopsis, etc. Although the sources are different, the molecular weight of HCYase is mostly around 170kDa, composed of four ho...

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Problems solved by technology

However, this method needs to be detected on a micro-fluorescence detector, which is not suitable for the application and promotion of clinical diagnosis and detection.

And this method is due to the FeCl 3 Considering that HCYase and fluorescein chromogenic agent DBPDA cannot be stored stably in an acidic environment, most of the kits using this method have three reagents, and the operation is cumbersome

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