Homocysteine diagnosis kit and homocysteine concentration measuring method

A technology of homocysteine ​​and diagnostic kits, applied in biological testing, color/spectral characteristic measurement, material inspection products, etc., can solve the problems of cumbersome operation, unsuitable for clinical diagnostic testing application and promotion, etc., and achieve low price , easy to detect, and conducive to the effect of promotion

Inactive Publication Date: 2016-12-07
ENZYMAKER LABCHANGZHOU CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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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

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  • Homocysteine diagnosis kit and homocysteine concentration measuring method
  • Homocysteine diagnosis kit and homocysteine concentration measuring method
  • Homocysteine diagnosis kit and homocysteine concentration measuring method

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Embodiment Construction

[0026] The present invention will now be further described in conjunction with specific examples, and the following examples are intended to illustrate the present invention rather than further limit the present invention.

[0027] 1. Preparation of a kit for detecting HCY content in human serum

[0028] R1 reagent: Add 10KU / L GLDH, 5mM TCEP, 0.5mM NADH, 15mMα-ketoglutaric acid, 0.01% Triton X-100, 0.05% sodium azide to 1L 100mM Tris-HCl buffer, stir evenly For the R1 reagent.

[0029] R2 reagent: Add 20KU / L HCYase, 0.05mM PLP, and 0.05% sodium azide to 1L of 100mM Tris-HCl buffer, and stir well to obtain R2 reagent.

[0030] Homocysteine ​​calibrator: In 3mM HCl solution, add 28μM HCY

[0031] Detection method:

[0032] Take Hitachi 7080 biochemical analyzer as an example: the measurement wavelength is 340nm, and the measurement method is kinetic method. The sample volume is 20uL, the R1 reagent is 220uL, and the R2 reagent is 30ul. The calibration method is linear measur...

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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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Application Information

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IPC IPC(8): G01N33/68G01N21/31G01N35/00
CPCG01N33/68G01N21/31G01N35/00
Inventor 周亚萍于海燕朱钰峰曹利民
Owner ENZYMAKER LABCHANGZHOU CO LTD
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