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Homocysteine diagnosing/measuring reagent (kit) and homocysteine concentration measuring method

A homocysteine ​​and reagent technology, applied in the field of medical testing and determination, can solve the problems of long test time, complicated operation, large variation of test data, etc.

Inactive Publication Date: 2010-06-23
SUZHOU ANJ BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Its characteristics are similar to those of high-performance liquid chromatography, but it has the disadvantages of complicated operation, long test time and large variation of test data.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] The homocysteine ​​diagnosis / measurement reagent of this embodiment is a single reagent, including:

[0048] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0049] Stabilizer 500mmol / L

[0050] Reduced coenzyme 0.25mmol / L

[0051] Homocysteine ​​desulfurase 9000U / L

[0052] 2-oxobutyryl synthase 8000U / L

[0053] Isocitrate dehydrogenase 10000U / L

[0054] Coenzyme A 3mmol / L

[0055] Oxidized ferredoxin 5mmol / L

[0056] 2-ketoglutaric acid 12mmol / L

[0057] After the reagents are all dissolved and prepared, they are divided into bottles and freeze-dried to make dry powder reagents; before use, add purified water and reconstitute for use.

[0058] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, initial absorbance 1.8±0.7, test main wavelength 340nm, test secondary wavelength 405nm, the volume ratio of the tested homocysteine ​​sample to the reagent is 1 / 25, the reaction direction is negative reaction (fall...

Embodiment 2

[0061] The homocysteine ​​diagnosis / measurement reagent of this embodiment is a dual reagent, including:

[0062] Reagent 1

[0063] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0064] Stabilizer 50mmol / L

[0065] Reduced coenzyme 0.25mmol / L

[0066] Coenzyme A 3mmol / L

[0067] Oxidized ferredoxin 5mmol / L

[0068] 2-ketoglutaric acid 12mmol / L

[0069] Reagent 2

[0070] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0071] Stabilizer 500mmol / L

[0072] Homocysteine ​​desulfurase 9000U / L

[0073] 2-oxobutyryl synthase 8000U / L

[0074] Isocitrate dehydrogenase 10000U / L

[0075] After the reagents are all dissolved and prepared, they are divided into bottles to make liquid double reagents, which can be used directly.

[0076] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, initial absorbance 1.8±0.7, test main wavelength 340nm, test secondary wavelength 405nm, tested homocysteine ​​sam...

Embodiment 3

[0079] The homocysteine ​​diagnosis / measurement reagents of this embodiment are three reagents, including:

[0080] Reagent 1

[0081] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0082] Stabilizer 50mmol / L

[0083] Reduced coenzyme 0.25mmol / L

[0084] Coenzyme A 3mmol / L

[0085] Oxidized ferredoxin 5mmol / L

[0086] 2-ketoglutaric acid 12mmol / L

[0087] Reagent 2

[0088] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0089] Stabilizer 500mmol / L

[0090] 2-oxobutyryl synthase 8000U / L

[0091] Isocitrate dehydrogenase 10000U / L

[0092] Reagent 3

[0093] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0094] Stabilizer 500mmol / L

[0095] Homocysteine ​​desulfurase 9000U / L

[0096] After all the reagents are dissolved and prepared, they are divided into bottles to make liquid three reagents, which can be used directly.

[0097] When measuring the concentration of homocysteine, set it on the automati...

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Abstract

The invention relates to a homocysteine diagnosing / measuring reagent (kit) using the technologies of an enzyme colorimetric method and an enzyme linkage method. At the same time, the invention also relates to a homocysteine concentration measuring method, the reagent composition and reagent ingredients, which belong to the technical field of medical detection and measurement. The reagent (kit) mainly comprises the following ingredients: a buffer solution, reduced form cozymase, cozymase A, oxidized form ferredoxin, 2-oxoglutarate, homocysteine desulfhydrase, 2-oxobutyryl synthetase, isocitrate dehydrogenase and stabilizing agents. The method has the following steps: mixing samples and the reagent by the certain volume percent for making the samples and the reagent take a series of enzymatic reaction, and then, placing reactants under an ultraviolet / visible light analyzer to detect the dropping degree of the light absorbancy in the position with the main wave length of 340 nm, so the concentration of the homocysteine can be measured and calculated.

Description

technical field [0001] The invention relates to a homocysteine ​​diagnosis / measurement reagent (box), and at the same time, the invention also relates to a method for measuring the homocysteine ​​concentration, which belongs to the technical field of medical examination and measurement. Background technique [0002] There are many methods for measuring homocysteine ​​in plasma, including high performance liquid chromatography (HPLC), amino acid analyzer assay, ion chromatography, enzyme-linked immunoassay (EIA), capillary gas chromatography-mass spectrometry, fluorescence Polarization immunoassay (FPIA), electrochemical method and enzymatic method, etc. [0003] 1. High-performance liquid chromatography (HPLC): It is a classic reference method, but it has the disadvantages of complicated operation, long test time, and large variation of test data. In clinical application, it is gradually adopted by enzyme-linked immunoassay, Fluorescence polarization analysis and enzymatic ...

Claims

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Application Information

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IPC IPC(8): G01N21/31G01N21/17G01N33/48C12Q1/26
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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