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Measuring method of homotype cysteine concentration and homotype cysteine diagnostic reagent kit

A homocysteine ​​and diagnostic kit technology, which is applied in the determination/inspection of microorganisms, chemical method analysis, biochemical equipment and methods, etc., can solve the problems of long test time, complicated operation and large test data variation, Achieve reliable test results, accurate test results, and simple methods

Inactive Publication Date: 2007-02-14
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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  • Measuring method of homotype cysteine concentration and homotype cysteine diagnostic reagent kit
  • Measuring method of homotype cysteine concentration and homotype cysteine diagnostic reagent kit
  • Measuring method of homotype cysteine concentration and homotype cysteine diagnostic reagent kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Embodiment one (single dose)

[0060] Prepare a homocysteine ​​diagnostic kit with the following ingredients and dosages:

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

[0062] Ammonium sulfate 50% (accounting for the total volume of the reagent),

[0063] NADH 0.25mmol / L,

[0064] Cystathionine β synthase 2000U / L,

[0065] Cystathionine β-lyase 2000U / L,

[0066] Glutamate dehydrogenase 50000U / L,

[0067] Lactate dehydrogenase 10000U / L,

[0068] Serine 50mmol / L,

[0069] α-ketoglutarate 16mmol / L.

[0070] 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.

[0071] 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 reage...

Embodiment 2

[0073] Embodiment two (two doses)

[0074] Prepare a homocysteine ​​diagnostic kit with the following ingredients and dosages:

[0075] Reagent I——

[0076] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L,

[0077] Glycerol 50% (accounting for the total volume of reagent I),

[0078] NADPH 0.25mmol / L,

[0079] Glutamate dehydrogenase 40000U / L,

[0080] Lactate dehydrogenase 15000U / L,

[0081] α-ketoglutarate 12mmol / L;

[0082] Reagent II——

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

[0084] Glycerol 50% (accounting for the total volume of reagent II),

[0085] Cystathionine β synthase 3000U / L,

[0086] Cystathionine β-lyase 4000U / L,

[0087] Serine 20mmol / L.

[0088] After the reagents are all dissolved and prepared, they are divided into bottles to make liquid double reagents, which can be used directly.

[0089] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, initial absor...

Embodiment 3

[0091] Embodiment three (three doses)

[0092] Prepare a homocysteine ​​diagnostic kit with the following ingredients and dosages:

[0093] Reagent I——

[0094] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L,

[0095] Propylene glycol 50% (accounting for reagent I total volume),

[0096] thio-NADH 0.25mmol / L,

[0097] α-ketoglutarate 10mmol / L;

[0098] Reagent II——

[0099] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L,

[0100] Propylene glycol 50% (accounting for total volume of reagent II),

[0101] Glutamate dehydrogenase 60000U / L,

[0102] Lactate dehydrogenase 20000U / L;

[0103] Reagent III——

[0104] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L,

[0105] Propylene glycol 50% (accounting for the total volume of reagent III),

[0106] Cystathionine β synthase 4000U / L,

[0107] Cystathionine β-lyase 4000U / L,

[0108] Serine 20mmol / L.

[0109] After all the reagents are dissolved and prepared, they a...

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PUM

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Abstract

A method for determining concentration of homotype cysteine includes synthesizing cystathionine B synthase, homotype cysteine and serine to be cystathionine; cracking cystathionine by cystathionine B lyase to change it back to be homotype cysteine and to generate ammonia and pyruvic acid; using some chemicals to act on ammonia to let reduction coenzyme be oxidized to be oxidation coenzyme for determining out absorbance dropping speed of reduction coenzyme at 340nm then estimating out concentration size of homotyp cysteine. Its diagnosis kit can be prepared in double dosages for decreasing cross influence of various compositions.

Description

technical field [0001] The invention relates to the detection of the concentration of homocysteine ​​in the field of medicine, in particular to a method for determining the concentration of homocysteine ​​by using enzyme cycle amplification method, enzyme colorimetric method and enzyme-linked method, and the homocysteine ​​prepared thereby. The cysteine ​​diagnostic kit belongs to the technical field of homocysteine ​​concentration analysis and detection. 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 t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31G01N21/25G01N31/00C12Q1/00
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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