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Method and measurement kit for measuring homocysteine by a cystathionine beta-synthase, a cystathionine beta-catabolic enzyme, and coupled pyruvic oxidase

A technology of homocysteine ​​and detection kit, applied in the field of laboratory medicine, can solve the problem of low sensitivity and so on

Pending Publication Date: 2019-10-18
浙江曼薇尼生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the low content of homocysteine ​​in the human body, the sensitivity of the method used in the conventional laboratory is not high. In recent years, the cyclic enzyme method has been invented

Method used

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  • Method and measurement kit for measuring homocysteine by a cystathionine beta-synthase, a cystathionine beta-catabolic enzyme, and coupled pyruvic oxidase
  • Method and measurement kit for measuring homocysteine by a cystathionine beta-synthase, a cystathionine beta-catabolic enzyme, and coupled pyruvic oxidase
  • Method and measurement kit for measuring homocysteine by a cystathionine beta-synthase, a cystathionine beta-catabolic enzyme, and coupled pyruvic oxidase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Reagent one:

[0031]

[0032] Reagent 2:

[0033]

[0034] TOOS: N-ethyl-N-(2-hydroxy-3-propanesulfo)-m-toluidine

[0035] Reagent 1: Reagent 2 is 2:1, rate method, positive reaction, wavelength: 546nm, tested sample: Reagent 1: Reagent 2 is 3.0: 180: 90, temperature 37°C, serum and reagent 1 react for 5 minutes, add reagent Second, delay for 1 minute, and detect the absorbance change rate for 2 minutes.

Embodiment 2

[0037] Reagent one

[0038]

[0039]

[0040] Reagent 2:

[0041]

[0042] Reagent 1: Reagent 2 is 3:1, rate method, positive reaction, wavelength: 510nm, tested sample: Reagent 1: Reagent 2 is 2.8: 210: 70, temperature 37°C, serum and reagent 1 react for 5 minutes, add reagent Second, delay for 1 minute, and detect the absorbance change rate for 2 minutes.

Embodiment 3

[0044] Reagent one:

[0045]

[0046]

[0047] Reagent 2:

[0048]

[0049] ADPS: Sodium N-ethyl-N-(3-sulfopropyl)-3-methoxyanilide

[0050] Reagent 1: Reagent 2 is 4:1, rate method, positive reaction, wavelength: 546nm, tested sample: Reagent 1: Reagent 2 is 2.6: 240: 60, temperature 37°C, serum and reagent 1 react for 5 minutes, add reagent Second, delay for 1 minute, and detect the absorbance change rate for 2 minutes.

[0051] Taking Example 3 as an example, the detection data are as follows:

[0052] The linearity result of the detection kit is

[0053]

[0054] Linear equation: y=0.975x+0.613, R 2 = 0.999. See attached figure 1 .

[0055] Detect the purchased quality control product (8.7~12.2~15.7mmol / L), repeat the test 20 times, the results are as follows:

[0056]

[0057] SD = 0.1598, CV = 1.323%.

[0058] Using the detection kit (Trinder method) and the purchased homocysteine ​​determination kit (ultraviolet method) to detect 50 clinical se...

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Abstract

The invention relates to a method for measuring homocysteine by a cystathionine beta-synthase, a cystathionine beta-catabolic enzyme, and coupled pyruvic oxidase. According to the principles: with tris(2-carboxyethyl)phosphorus hydrogen chloride, oxidized homocysteine is converted into free homocysteine; L-cystathionine is generated under catalysis of cystathionine beta-synthetase and alpha-amino-beta-hydroxyrpropionic acid reaction; homocysteine, ammonia and pyruvic acid are generated under the catalysis of cystathionine beta-clastic enzyme on the L-cystathionine; hydrogen peroxide is generated under the catalysis of pyruvic oxidase on the pyruvic acid; and then Trinder reaction color developing is performed. The homocysteine produced during the reaction makes reaction again to form a cyclic reaction. Besides, a homocysteine measurement kit can be made.

Description

technical field [0001] The invention belongs to the field of laboratory medicine, and relates to a method for measuring homocysteine ​​and a test kit. Background technique [0002] Homocysteine ​​is a sulfur-containing amino acid and an important intermediate product in the metabolism of methionine. The detection of homocysteine ​​is mainly used clinically as an important indicator of cardiovascular diseases, especially coronary atherosclerosis and myocardial infarction, and its concentration is directly proportional to the risk of the disease. Homocysteinemia is closely related to cardiovascular diseases, and elevated concentrations can damage coronary arteries and other blood vessels, eventually causing atherosclerosis and other cardiovascular diseases. Vitamin B 6 , B 12 And folic acid, by participating in homocysteine ​​metabolism, regulates and reduces homocysteine, thereby reducing the risk of cardiovascular diseases. Homocysteine ​​is an independent risk factor fo...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N21/78
CPCG01N33/6815G01N21/78
Inventor 李丰盛倪建芬车桃李
Owner 浙江曼薇尼生物科技有限公司
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