Homotypic cysteine measuring method and its reagent

A homocysteine, cysteinase technology, applied in biological testing, material analysis by observing the impact on chemical indicators, material testing products, etc. Can not apply clinical large flow automatic analysis and other problems

Active Publication Date: 2005-01-05
北京安百胜诊断科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The common features of the above methods are that special measuring instruments are required, the operation is complicated, the test takes a long time, it cannot be applied to the automatic analysis of clinical large flow, and it is expensive
[0004] Due to the low content of homocysteine ​​in the sample, conventional clinical enzymology assay methods cannot achieve the required assay sensitivity, which limits the adoption of conventional methods
In terms of technology for the determination of homocysteine ​​by enzymatic methods,

Method used

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  • Homotypic cysteine measuring method and its reagent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Embodiment 1: The hydrogen peroxide chromogenic reaction (Trinder's reaction) produced by the reagent system is determined by using an enzyme cycle reaction consisting of homocysteine ​​methyltransferase and adenosyl homocysteine.

[0020] Reagent components

[0021] AdoHcyase

[0022] Reagent components

[0023]Example 1 Reagent 1 formula is used to reduce oxidized homocysteine ​​to eliminate the interference of adenosine in the sample, and the complete enzyme cycle assay reagent of reagent 1 is used to eliminate side reactions. When measuring samples, the fixed time method is adopted, and the ratio of reagents to samples is not fixed, but it should be consistent within the same measurement batch. For example, the ratio of reagent 1: sample: reagent 2 is 300: 20: 100, the temperature is 37°C, and the measurement wavelength is 540 nm. After adding the sample or calibrator to reagent 1, incubate at the measurement temperature for 300 seconds ...

Embodiment 2

[0024] Example 2: Using an enzyme cycle reaction consisting of homocysteine ​​methyltransferase and adenosylhomocysteinase, the rate of oxidation of reduced coenzyme within a fixed period of time was determined by ammonia reagent method.

[0025] Reagent components

[0026] DTT

[0027] Reagent components

[0028] Example 2 Reagent 1 formula is used to reduce oxidized homocysteine, and Reagent 2 and Reagent 1 are combined to form a complete enzyme cycle assay reagent. When measuring samples, the fixed time method is adopted, and the ratio of reagents to samples is not fixed, but it should be consistent within the same measurement batch. For example, the ratio of reagent 1: sample: reagent 2 is 200: 25: 50, the temperature is 37°C, and the measurement wavelength is 340 nm. After adding sample or calibrator to reagent 1, incubate at the measurement temperature for 300 seconds, then add reagent 2, delay for 60 to 120 seconds, and eliminate The interf...

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PUM

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Abstract

The invention relates to an enzymology measuring method and reagent with circular increment technology, applied to measure the content of homeotypic cysteine in the measured sample. The character is: with a circular increment technology, increases the measurement sensitivity, thus it can carries on automatic measurement to the homeotypic cysteine in the sample liquid like normal enzymology diagnosing agent. The homeotypic cysteine in the sample liquid reacts with the S-adenosine-L-methionine repeatedly and circularly under the effect of homeotypic cysteine methyl kinase and adenosine homeotypic cysteine, generates the adenosine, the speed of the adenosine generation is in direct proportion to the homeotypic cysteine in the sample, through the generation speed, the aim to measure the content of the homeotypic cysteine in sample can be achieved.

Description

Technical field: [0001] The invention relates to a method for determining homocysteine ​​and its reagents Background technique: [0002] Homocysteine ​​(Homocysteine) is a sulfur-containing amino acid that is produced in cells by demethylation of methionine. Recent studies have shown that homocysteine ​​produces superoxide and peroxide damages vascular endothelial cells, changes the function of coagulation factors, increases the tendency of thrombosis, promotes atherosclerosis and thrombosis, and increases the incidence of cardiovascular diseases and Mortality increases, so measuring the concentration of homocysteine ​​in blood is of great clinical significance. When homocysteine ​​accumulates in cells and enters the blood circulation, most of it exists in the oxidized form and binds to proteins, while the reduced form of homocysteine ​​accounts for only 1% in the blood. To determine the content of total homocysteine ​​in blood, it is generally necessary to use a reducing ...

Claims

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Application Information

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IPC IPC(8): G01N21/78G01N33/68
Inventor 王学忠
Owner 北京安百胜诊断科技有限公司
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