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Homocysteine diagnostic kit and method for measuring homocysteine concentration

A technology of homocysteine ​​and reagents, which is applied in the field of medical inspection and measurement, and can solve the problems of long test time, large variation of test data, complicated operation, etc.

Inactive Publication Date: 2009-12-23
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 its disadvantages are 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

[0046] The homocysteine ​​diagnostic reagent of this embodiment is a single reagent, including:

[0047] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0048] Stabilizer 500mmol / L

[0049] Reduced coenzyme 0.25mmol / L

[0050] Homocysteine-α, γ-lyase 6000U / L

[0051] Alanine dehydrogenase 8000U / L

[0052] Glycine oxidase 10000U / L

[0053] NADH peroxidase 10000U / L

[0054] Pyruvate 15mmol / L

[0055] 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.

[0056] 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 (falling reaction), the delay time is about 0 minutes, and the detectio...

Embodiment 2

[0059] The homocysteine ​​diagnostic reagent of this embodiment is a dual reagent, including:

[0060] Reagent 1

[0061] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0062] Stabilizer 50mmol / L

[0063] Reduced coenzyme 0.25mmol / L

[0064] Pyruvate 15mmol / L

[0065] Reagent 2

[0066] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0067] Stabilizer 500mmol / L

[0068] Homocysteine-α, γ-lyase 6000U / L

[0069] Alanine dehydrogenase 8000U / L

[0070] Glycine oxidase 10000U / L

[0071] NADH peroxidase 10000U / L

[0072] After the reagents are all dissolved and prepared, they are divided into bottles to make liquid double reagents, which can be used directly.

[0073] 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 ​​sample and reagent 1, reagent 2 The volume ratio is 2 / 20 / 5, the reac...

Embodiment 3

[0076] The homocysteine ​​diagnostic reagent of this embodiment is three reagents, including:

[0077] Reagent 1

[0078] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0079] Stabilizer 50mmol / L

[0080] Reduced coenzyme 0.25mmol / L

[0081] Pyruvate 15mmol / L

[0082] Reagent 2

[0083] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0084] Stabilizer 500mmol / L

[0085] Alanine dehydrogenase 8000U / L

[0086] Glycine oxidase 10000U / L

[0087] NADH peroxidase 10000U / L

[0088] Reagent 3

[0089] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0090] Stabilizer 500mmol / L

[0091] Homocysteine-α, γ-lyase 6000U / L

[0092] After all the reagents are dissolved and prepared, they are divided into bottles to make liquid three reagents, which can be used directly.

[0093] When measuring the concentration of homocysteine, set it on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, ...

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Abstract

The invention relates to a homocysteine diagnostic kit utilizing technologies of an enzymatic recycling method, an ELISA multiplication method, an enzymatic-colorimetric method and an enzyme-link method, also relates to a method and a principle of measuring homocysteine concentration and compositions and components of reagents, and belongs to the technical field of testing and measuring of medical science. The kit mainly comprises the following compositions: buffer solution, reduced coenzyme, pyruvic acid, homocysteine-alpha, gamma-lyase, alanine dehydrogenase, glycine oxidase, NADH peroxidase, and stabilizer; samples are mixed with the reagents in certain volume ratio to perform a series of enzymatic reactions; then reactants are placed under a UV / visible analyzer; and the descending speed of absorbance is tested at the position where dominant wave length is 340nm so as to measure and calculate the homocysteine concentration.

Description

technical field [0001] The invention relates to a homocysteine ​​diagnostic kit, and at the same time, the invention also relates to a method for measuring the concentration of homocysteine, which belongs to the technical field of medical examination and determination. Background technique [0002] There are many methods for measuring homocysteine ​​concentration 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] High-performance liquid chromatography (HPLC): It is a classic reference method, but its operation is relatively complicated, the test time is relatively long, and the test data varies greatly. It is gradually adopted by enzyme-linked immunoassay, fluorescence polarization analysis and enzymatically replaced. High-perf...

Claims

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Application Information

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IPC IPC(8): G01N21/31G01N21/25G01N33/68C12Q1/527C12Q1/26
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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