CN101609018A - Homotype semicystionl diagnostic kit and homocysteine method for measurement of concentration - Google Patents
Homotype semicystionl diagnostic kit and homocysteine method for measurement of concentration Download PDFInfo
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Abstract
The present invention relates to a kind of homotype semicystionl diagnostic kit that utilizes indirect enzyme cycle amplification method, enzymic colorimetric and enzyme-linked method technology, the invention still further relates to the method principle of measuring homocysteine concentration, the composition and the composition of reagent simultaneously, belong to medical test determination techniques field.Kit principal ingredient of the present invention comprises: damping fluid, 2-oxoglutaric acid, reduced coenzyme, homocysteine-alpha, gamma-lyases, glutamte dehydrogenase, dglutamic oxidase, peroxidase, the combination of reduced form chromogen and stabilizing agent; Pass through series of enzymatic reactions, the most colourless reduced form chromogen combination is oxidized to coloured dyestuff, thereby can pass through the visible light analysis instrument, at 400-700nm wavelength place, measure the height of dyestuff content, and then directly reflect the size of homocysteine concentration.
Description
Technical field
The present invention relates to a kind of homotype semicystionl diagnostic kit, the invention still further relates to the method for measuring homocysteine concentration simultaneously, belong to medical test determination techniques field.
Background technology
The method of measuring homocysteine in blood plasma concentration has a variety of, comprise high performance liquid chromatography (HPLC), amino-acid analyzer determination method, the chromatography of ions, enzyme-linked immunosorbent assay (EIA), capillary gas chromatography-mass spectrum, fluorescence polarization immunoassay (FPIA), electrochemical process and enzyme process etc.
High performance liquid chromatography (HPLC): be classical reference method, but its operation more complicated, test consuming time longly that the test figure variance ratio is bigger, is replaced by enzyme-linked immunosorbent assay, fluorescence polarization assay method and enzyme process gradually aspect clinical practice.High performance liquid chromatography: use earlier reductive agent to make that the homocysteine of form of ownership is transformed into the reduction form in the blood sample, with the fluorescer generation homocysteine-fluorescent material compound of deriving, the sample after will deriving carries out stratographic analysis then.Absorption affinity difference because of the relative different material of chromatographic stationary, therefore moving phase is also different with its order that elutes, according to this principle separately with the different material in the sample, detect the fluorescence intensity of homocysteine-fluorescent material compound under the wash-out with fluorescence detector, itself and standard items/interior target ratio are compared and calculate, just can measure the level of blood total homocysteine.
Enzyme-linked immunosorbent assay (EIA): be the common method of measuring homocysteine level clinically, operate more convenient, quick, good reproducibility, method is reliable and stable.Its shortcoming is most of operation or manual operations, and is more consuming time.Enzyme-linked immunosorbent assay fundamental analysis principle: use enzyme that the homocysteine of form of ownership in the blood sample is transformed into S-adenosine-L-homocysteine earlier, the monoclonal or the polyclonal antibody that add the anti-S-adenosine of enzyme target-L-homocysteine then, adopt the principle of competition combination, the standard items of varying level are combined with the enzyme labelled antibody competition, with chromogenic reagent with stop reagent and develop the color respectively and stop, produce the typical curve of homocysteine concentration and coloring intensity then.Sample is also handled by same steps as, just can find the concentration of its homocysteine on typical curve.
The chromatography of ions: be mainly used in experimental study, less clinically use, its fundamental analysis principle is a kind of of stratographic analysis, mainly is that its stationary phase adopts ion exchange material, homocysteine can be separated with other material according to its absorption affinity difference to different ions and measure.
Electrocapillary phoresis method: be mainly used in experimental study, the report in clinical use is arranged.Its characteristics are similar to efficient liquid-phase chromatography method, its operation more complicated, the test long and bigger shortcoming of test figure variance ratio consuming time.The fundamental analysis principle: use earlier reductive agent to make that the homocysteine of form of ownership is transformed into the reduction form in the blood sample, with the fluorescent reagent generation homocysteine-fluorescent material compound of deriving, the sample after will deriving carries out the electrocapillary phoresis analysis then.Sample is positioned in the high-voltage electric field about 10 kilovolts, because the electrically charged difference of various materials, its isoelectric point is also inequality, therefore its migration velocity in electric field is also just different, homocysteine and other material in the sample can be separated according to this principle, detect the fluorescence intensity of homocysteine-fluorescent material compound again with fluorescence detector, itself and standard items/interior target ratio are compared and calculate, just can measure the level of total homocysteine.
Fluorescence polarization method: be the reasonable method of measuring homocysteine level clinically, this method full automation instrumental analysis, have fast, accurately, advantage easily.Its fundamental analysis principle: the fluorescence polarization intensity of the homocysteine that free homocysteine and anti-monoclonal antibody combine in the sample is different, sample, standard items are combined with the labeling antibody competition, adopt the principle of competing combination to produce the typical curve of homocysteine concentration and fluorescence polarization intensity, on typical curve, just can find the concentration of its homocysteine.
Enzyme process: the method for testing of coming out newly developed in response to the market demand, at present domestic have a multinomial patented claim, and CN98807531.8 utilizes homocysteine to come homocysteine content in the measuring samples; CN200410016789.6 utilizes the cyclic amplification effect of HcyMetase (E.C.2.1.1.10) and S-Adenosylhomocysteine synthase (E.C.3.3.1.1), add adenosine deaminase, purine nucleoside phosphorylase, xanthine oxidase, peroxidase etc., or chromogenic reactions such as adenosine deaminase, glutamte dehydrogenase are measured homocysteine content; CN200510053210.8 utilizes the L-methionine Gamma-catenase to act on and produces fluorescence with fluorescent Compound D MPD2HCl effect again after homocysteine makes it to produce sulfuretted hydrogen, this method is used the test of full-automatic fluorescence analyser, have fast, accurately, advantage easily.Its fundamental analysis principle: with the genetic recombination enzyme homocysteine is decomposed, the sulfuretted hydrogen product that is formed forms measurable fluorescent chemicals with fluorescent color-developing agent generation chemical reaction again.
Summary of the invention
The technical problem to be solved in the present invention is: propose a kind of indirect enzyme cycle amplification method (IndirectEnzymatic Recycling Method) of utilizing, enzymic colorimetric (Enzymatic Colorimetric Method) and enzyme-linked method (Couple Reaction) technology, metering generates the rising in 400-700nm wavelength place absorbance of indoleamine chromogen (Indamine dye) or quinone-imine chromogen (Quioneimine dye) cause by integrated enzyme reaction, measured the method for homocysteine concentration, simultaneously, the present invention also will provide in order to realize the homotype semicystionl diagnostic kit of this method, adopt this reagent not only can be visible light analysis instrument or half, carry out the homocysteine concentration determination on the automatic clinical chemistry analyzer, and finding speed is fast, sensitivity is big, the accuracy height, thereby can obtain practical applying.
Homocysteine method for measurement of concentration principle of the present invention is as follows:
Homocysteine+water
Homocysteine-alpha, gamma-lyasesSulfuretted hydrogen+ammonia+
The alpha-oxo-butyric acid
Ammonia+2-oxoglutaric acid+reduced coenzyme
Glutamte dehydrogenaseGlutamic acid+coenzyme+water
Glutamic acid+oxygen+water
Dglutamic oxidaseAmmonia+2-oxoglutaric acid+hydrogen peroxide
Hydrogen peroxide+reduced form chromogen combination
PeroxidaseThe indoleamine chromogen or
Quinone-imine chromogen+water
This method is used homocysteine-alpha, gamma-lyases (homocysteine desulfhydrase; EC4.4.1.2) (idol) connection glutamte dehydrogenase (Glutamate Dehydrogenase; EC 1.4.1.2; EC 1.4.1.3; EC 1.4.1.4), dglutamic oxidase (Glutamate oxidase; EC 1.4.3.7; EC 1.4.3.11) and peroxidase (peroxidase; EC 1.11.1.7) enzyme ' s reaction speeding colourimetry.Homocysteine-alpha, gamma-lyases is as the effect enzyme, and function is for to separate generation ammonia with homocysteine.Glutamte dehydrogenase is also as the effect enzyme, and the enzymatic catalysis of glutamte dehydrogenase makes ammonia produce glutamic acid; Dglutamic oxidase is as cyclophorase: dglutamic oxidase becomes glutamic acid again ammonia more once more, and it is recycling that the ammonia that is produced by homocysteine constantly is repeated, and therefore continuously produces hydrogen peroxide.Peroxidase (peroxidase; EC 1.11.1.7) as the colour developing enzyme, by with the effect of hydrogen peroxide, make colourless reduced form chromogen combination be oxidized to coloured dyestuff, thereby can pass through the visible light analysis instrument, measure the light absorption size speed of dyestuff-indoleamine chromogen (Indamine dye) or quinone-imine chromogen (Quioneimine dye)-content height at 400-700nm (difference according to the combination of reduced form chromogen is decided) wavelength place, draw homocysteine concentration measurement result.
Experiment shows, takes all factors into consideration from the accuracy of measurement result and economy two aspects of preparation cost, and the homotype semicystionl diagnostic kit of the present invention of following composition relation is comparatively desirable:
Damping fluid 100mmol/L
Stabilizing agent 500mmol/L
Peroxidase 30000U/L
Homocysteine-alpha, gamma-lyases 10000U/L
Glutamte dehydrogenase 12000U/L
Dglutamic oxidase 12000U/L
2-oxoglutaric acid 5mmol/L
Reduced coenzyme 1mmol/L
Reduced form chromogen combination 0.1---20mmol/L
Described reduced form chromogen combination (Chromogen) can be any one combinations of pairs in following 28 combinations:
3-methyl-2-2-thiobenzimide hydrazone
Carbolic acid
3-methyl-2-2-thiobenzimide hydrazone
N-ethyl-N-(3-thiopropyl)-m-thialdine amine
3-methyl-2-2-thiobenzimide hydrazone
N, the two ethyls of N--m-toluidine
3-methyl-2-2-thiobenzimide hydrazone
2, the two chlorine carbolic acids of 4-
3-methyl-2-2-thiobenzimide hydrazone
2,4,6-three bromo-3-hydroxyl-benzene sulfonic acid
3-methyl-2-2-thiobenzimide hydrazone
3, the two chlorine carbolic acid sulfonic acid of 5-
3-methyl-2-2-thiobenzimide hydrazone
3, the two chloro-2-hydroxyl-benzene sulfonic acids of 5-
3-methyl-2-2-thiobenzimide hydrazone
N-ethyl-N-(2-Hydroxyalkyl base-3-thiopropyl)-m-toluidine sodium salt
3-methyl-2-2-thiobenzimide hydrazone
Sa Xiu Hydroxyalkyl yl benzoic acid
3-methyl-2-2-thiobenzimide hydrazone
Two methylanilines
3-methyl-2-2-thiobenzimide hydrazone
N-ethyl-N-(2-Hydroxyalkyl base-3-thiopropyl)-m-toluidine
3-methyl-2-2-thiobenzimide hydrazone
2,2 '-AZINO-two (3-ethylbenzene thiazole-6-sulfonic acid)
3-methyl-2-2-thiobenzimide hydrazone
4-Hydroxyalkyl base-3-methoxy benzoic acid
3-methyl-2-2-thiobenzimide hydrazone
3-methyl-ethyl-Hydroxyalkyl base aniline
The amino anti-arsenic of 4-
Carbolic acid
The amino anti-arsenic of 4-
N-ethyl-N-(3-thiopropyl)-m-thialdine amine
The amino anti-arsenic of 4-
N, the two ethyls of N--m-toluidine
The amino anti-arsenic of 4-
2, the two chlorine carbolic acids of 4-
The amino anti-arsenic of 4-
2,4,6-three bromo-3-hydroxyl-benzene sulfonic acid
The amino anti-arsenic of 4-
3, the two chlorine carbolic acid sulfonic acid of 5-
The amino anti-arsenic of 4-
3, the two chloro-2-hydroxyl-benzene sulfonic acids of 5-
The amino anti-arsenic of 4-
N-ethyl-N-(2-Hydroxyalkyl base-3-thiopropyl)-m-toluidine sodium salt
The amino anti-arsenic of 4-
Sa Xiu Hydroxyalkyl yl benzoic acid
The amino anti-arsenic of 4-
Two methylanilines
The amino anti-arsenic of 4-
N-ethyl-N-(2-Hydroxyalkyl base-3-thiopropyl)-m-toluidine
The amino anti-arsenic of 4-
2,2 '-AZINO-two (3-ethylbenzene thiazole-6-sulfonic acid)
The amino anti-arsenic of 4-
4-Hydroxyalkyl base-3-methoxy benzoic acid
The amino anti-arsenic of 4-
3-methyl-ethyl-Hydroxyalkyl base aniline
Homotype semicystionl diagnostic kit of the present invention can be single agent, comprising:
Damping fluid, stabilizing agent, homocysteine-alpha, gamma-lyases, glutamte dehydrogenase, dglutamic oxidase, 2-oxoglutaric acid, reduced coenzyme, peroxidase, the combination of reduced form chromogen.Kit can be a dry powder, and use the back that is dissolved in water before use; Also can be mixed with liquid reagent, directly use.
Also above-mentioned single agent reagent can be made into following pair of agent reagent:
Reagent 1
Damping fluid, stabilizing agent, the combination of reduced form chromogen, 2-oxoglutaric acid, reduced coenzyme.
Reagent 2
Damping fluid, stabilizing agent, peroxidase, the combination of reduced form chromogen, homocysteine-alpha, gamma-lyases, glutamte dehydrogenase, dglutamic oxidase.
Peroxidase, the combination of reduced form chromogen, homocysteine-alpha, gamma-lyases, glutamte dehydrogenase, dglutamic oxidase, 2-oxoglutaric acid, the position of reduced coenzyme in reagent 1 or reagent 2 can not limit.Kit can be a dry powder, and use the back that is dissolved in water before use; Also can be mixed with liquid reagent, directly use.
Also above-mentioned single agent reagent can be made into following three doses of reagent:
Reagent 1
Damping fluid, stabilizing agent, the combination of reduced form chromogen, 2-oxoglutaric acid, reduced coenzyme.
Reagent 2
Damping fluid, stabilizing agent, the combination of reduced form chromogen, glutamte dehydrogenase, dglutamic oxidase, peroxidase.
Reagent 3
Damping fluid, stabilizing agent, homocysteine-alpha, gamma-lyases.
Peroxidase, the combination of reduced form chromogen, homocysteine-alpha, gamma-lyases, glutamte dehydrogenase, dglutamic oxidase, 2-oxoglutaric acid, the position of reduced coenzyme in reagent 1, reagent 2 or reagent 3 can not limit.Kit can be a dry powder, and use the back that is dissolved in water before use; Also can be mixed with liquid reagent, directly use.
Embodiment
The present invention is further illustrated below in conjunction with examples of implementation.
Embodiment one
The homocysteine diagnosis reagent of present embodiment is single reagent, comprising:
Three (ethyloic) aminomethane-hydrochloride buffer 100mmol/L
Stabilizing agent 500mmol/L
Homocysteine-alpha, gamma-lyases 10000U/L
Glutamte dehydrogenase 12000U/L
Dglutamic oxidase 12000U/L
2-oxoglutaric acid 5mmol/L
Reduced coenzyme 1mmol/L
Peroxidase 30000U/L
The amino anti-arsenic 2mmol/L of 4-
Carbolic acid 10mmol/L
Reagent divides the bottle of packing into after all dissolving and preparing, and carries out freeze drying, makes powdered reagent; Before the use, add pure water, use after redissolving.
On automatic clinical chemistry analyzer, set: 37 ℃ of temperature, 10 minutes reaction time, test predominant wavelength 505nm, test commplementary wave length 600nm, the volume ratio of tested homocysteine sample and reagent is 1/25, and the Direction of Reaction is positive reaction (reaction of rising), and detection method is a rate method, about about 1 minute of time delay is about 2 minutes detection times.
After adding sample and reagent, make them mixed and have reaction, reactant places under the Biochemical Analyzer the most at last, detects the speed that predominant wavelength 505nm absorbance rises, thereby calculates the concentration of homocysteine.
Embodiment two
The homocysteine diagnosis reagent of present embodiment is double reagent, comprising:
Reagent 1
Three (ethyloic) aminomethane-hydrochloride buffer 100mmol/L
Stabilizing agent 50mmol/L
2-oxoglutaric acid 5mmol/L
Reduced coenzyme 1mmol/L
2,4,6-three bromo-3-hydroxyl-benzene sulfonic acid 0.2mmol/L
Reagent 2
Three (ethyloic) aminomethane-hydrochloride buffer 100mmol/L
Stabilizing agent 500mmol/L
Homocysteine-alpha, gamma-lyases 10000U/L
Glutamte dehydrogenase 12000U/L
Dglutamic oxidase 12000U/L
Peroxidase 30000U/L
The amino anti-arsenic 0.6mmol/L of 4-
Reagent divides the bottle of packing into after all dissolving and preparing, and makes liquid double reagent, can directly use.
On automatic clinical chemistry analyzer, set: 37 ℃ of temperature, 10 minutes reaction time, test predominant wavelength 546nm, test commplementary wave length 600nm, the volume ratio of tested homocysteine sample and reagent is 1/20, and the Direction of Reaction is positive reaction (reaction of rising), and detection method is a rate method, about about 1 minute of time delay is about 2 minutes detection times.
After adding sample and reagent, make them mixed and have reaction, reactant places under the Biochemical Analyzer the most at last, detects the speed that predominant wavelength 546nm absorbance rises, thereby calculates the concentration of homocysteine.
Embodiment three
The homocysteine diagnosis reagent of present embodiment is three reagent, comprising:
Reagent 1
Three (ethyloic) aminomethane-hydrochloride buffer 100mmol/L
Stabilizing agent 50mmol/L
2-oxoglutaric acid 5mmol/L
Reduced coenzyme 1mmol/L
3-methyl-2-2-thiobenzimide hydrazone 0.6mmol/L
Reagent 2
Three (ethyloic) aminomethane-hydrochloride buffer 100mmol/L
Stabilizing agent 500mmol/L
Glutamte dehydrogenase 12000U/L
Dglutamic oxidase 12000U/L
Peroxidase 30000U/L
Two methylaniline 2mmol/L
Reagent 3
Three (ethyloic) aminomethane-hydrochloride buffer 100mmol/L
Stabilizing agent 500mmol/L
Homocysteine-alpha, gamma-lyases 10000U/L
Reagent divides the bottle of packing into after all dissolving and preparing, and makes liquid three reagent, can directly use.
On automatic clinical chemistry analyzer, set: 37 ℃ of temperature, 10 minutes reaction time, test predominant wavelength 578nm, test commplementary wave length 660nm, the volume ratio of tested homocysteine sample and reagent is 1/30, and the Direction of Reaction is positive reaction (reaction of rising), and detection method is a rate method, about about 1 minute of time delay is about 2 minutes detection times.
After adding sample and reagent, make them mixed and have reaction, reactant places under the Biochemical Analyzer the most at last, detects the speed that predominant wavelength 578nm absorbance rises, thereby calculates the concentration of homocysteine.
The applicant adopts other various reduced form chromogens combinations of putting down in writing in the above summary of the invention all can reach purpose of the present invention through experimental verification, in view of situation such as determination step and above embodiment roughly the same, do not separately enumerate.
In a word, experimental results show that: adopt assay method of the present invention can draw required measurement result by general biochemical analyzer fully---the blank reagent absorbance changes (Δ A/min)≤0.0007; Absorbance time response curve should be the rising curve until terminal point; Reagent can be surveyed effectively, and (R 〉=0.99) linear range can reach 0.7mmol/L; The inaccuracy of reagent test, its relative deviation be no more than ± and 5%; The coefficient of variation (CV)≤2% of the precision of reagent test (repeatability); The sensitivity of reagent can reach 0.6 ± 0.3 Δ A/mmol/L; Reagent is preserved down at 2-8 ℃, and activity can be stablized 1 year;---the present invention is highly sensitive, degree of accuracy good, the linear range broadness, and stationary phase is long, is enough to easy to utilize.
Claims (7)
1. the homocysteine method for measurement of concentration of an indirect enzyme cycle amplification method, enzymic colorimetric and enzyme-linked method technology, its method principle is as follows:
Homocysteine+water
Homocysteine-alpha, gamma-lyasesSulfuretted hydrogen+ammonia+
The alpha-oxo-butyric acid
Ammonia+2-oxoglutaric acid+reduced coenzyme
Glutamte dehydrogenaseGlutamic acid+coenzyme+water
Glutamic acid+oxygen+water
Dglutamic oxidaseAmmonia+2-oxoglutaric acid+hydrogen peroxide
Hydrogen peroxide+reduced form chromogen combination
PeroxidaseThe indoleamine chromogen or
Quinone-imine chromogen+water
The end reaction thing is placed under the visible light analysis instrument, detect the light absorption speed that indoleamine chromogen or quinone-imine chromogen content height are directly reflected in the 400-700nm place, draw homocysteine concentration measurement result.
2. homotype semicystionl diagnostic kit, principal ingredient comprises:
Damping fluid 20---500mmol/L
Stabilizing agent 1---4000mmol/L
Peroxidase 500---80000U/L
Homocysteine-alpha, gamma-lyases 1000---80000U/L
Glutamte dehydrogenase 1000---80000U/L
Dglutamic oxidase 1000---80000U/L
2-oxoglutaric acid 1---50mmol/L
Reduced coenzyme 1---50mmol/L
Reduced form chromogen combination 0.1---20mmol/L
The concentration of reagent composition not necessarily is only limited to above-mentioned scope; Effect is better in this scope, and outside this scope, reagent still can reagentia.
It is characterized in that: kit can be a dry powder, and use the back that is dissolved in water before use; Also can be mixed with liquid reagent, directly use.
3. according to the described homotype semicystionl diagnostic kit of claim 2, it is characterized in that: form single agent reagent by damping fluid, stabilizing agent, homocysteine-alpha, gamma-lyases, glutamte dehydrogenase, dglutamic oxidase, 2-oxoglutaric acid, reduced coenzyme, peroxidase, reduced form chromogen.
4. according to the described homotype semicystionl diagnostic kit of claim 2, it is characterized in that: form two agent reagent by damping fluid, stabilizing agent, homocysteine-alpha, gamma-lyases, glutamte dehydrogenase, dglutamic oxidase, 2-oxoglutaric acid, reduced coenzyme, peroxidase, reduced form chromogen; Reagent 1 is made up of damping fluid, stabilizing agent, 2-oxoglutaric acid, reduced coenzyme, reduced form chromogen; Reagent 2 is made up of damping fluid, stabilizing agent, homocysteine-alpha, gamma-lyases, glutamte dehydrogenase, dglutamic oxidase, peroxidase, reduced form chromogen.Peroxidase, the combination of reduced form chromogen, homocysteine-alpha, gamma-lyases, glutamte dehydrogenase, dglutamic oxidase, 2-oxoglutaric acid, the position of reduced coenzyme in reagent 1 or reagent 2 can not limit.
5. according to the described homotype semicystionl diagnostic kit of claim 2, it is characterized in that: form multi-agent reagent by damping fluid, stabilizing agent, homocysteine-alpha, gamma-lyases, glutamte dehydrogenase, dglutamic oxidase, 2-oxoglutaric acid, reduced coenzyme, peroxidase, reduced form chromogen; Reagent 1 is made up of damping fluid, stabilizing agent, 2-oxoglutaric acid, reduced coenzyme, reduced form chromogen; Reagent 2 is made up of damping fluid, stabilizing agent, glutamte dehydrogenase, dglutamic oxidase, peroxidase, reduced form chromogen; Reagent 3 is made up of damping fluid, stabilizing agent, homocysteine-alpha, gamma-lyases.Peroxidase, reduced form chromogen combination homocysteine-alpha, gamma-lyases, glutamte dehydrogenase, dglutamic oxidase, 2-oxoglutaric acid, the position of reduced coenzyme in reagent 1, reagent 2 or reagent 3 can not limit.
6. according to the described homotype semicystionl diagnostic kit of claim 2, it is characterized in that: also comprise stabilizing agent 1-4000mmol/L or 0.1%-100% volume ratio.Described stabilizing agent is: ammonium sulfate (AmmoniaSulfate), glycerine (Glycerol), propylene glycol (Propylene Glycol), ethylene glycol (Ethylene glycol) and at least one of the preservatives.
7. according to the described homotype semicystionl diagnostic kit of claim 2, it is characterized in that: described reduced form chromogen combination can be any combinations of pairs in following 28 combinations:
3-methyl-2-2-thiobenzimide hydrazone
Carbolic acid
3-methyl-2-2-thiobenzimide hydrazone
N-ethyl-N-(3-thiopropyl)-m-thialdine amine
3-methyl-2-2-thiobenzimide hydrazone
N, the two ethyls of N--m-toluidine
3-methyl-2-2-thiobenzimide hydrazone
2, the two chlorine carbolic acids of 4-
3-methyl-2-2-thiobenzimide hydrazone
2,4,6-three bromo-3-hydroxyl-benzene sulfonic acid
3-methyl-2-2-thiobenzimide hydrazone
3, the two chlorine carbolic acid sulfonic acid of 5-
3-methyl-2-2-thiobenzimide hydrazone
3, the two chloro-2-hydroxyl-benzene sulfonic acids of 5-
3-methyl-2-2-thiobenzimide hydrazone
N-ethyl-N-(2-Hydroxyalkyl base-3-thiopropyl)-m-toluidine sodium salt
3-methyl-2-2-thiobenzimide hydrazone
Sa Xiu Hydroxyalkyl yl benzoic acid
3-methyl-2-2-thiobenzimide hydrazone
Two methylanilines
3-methyl-2-2-thiobenzimide hydrazone
N-ethyl-N-(2-Hydroxyalkyl base-3-thiopropyl)-m-toluidine
3-methyl-2-2-thiobenzimide hydrazone
2,2 '-AZINO-two (3-ethylbenzene thiazole-6-sulfonic acid)
3-methyl-2-2-thiobenzimide hydrazone
4-Hydroxyalkyl base-3-methoxy benzoic acid
3-methyl-2-2-thiobenzimide hydrazone
3-methyl-ethyl-Hydroxyalkyl base aniline
The amino anti-arsenic of 4-
Carbolic acid
The amino anti-arsenic of 4-
N-ethyl-N-(3-thiopropyl)-m-thialdine amine
The amino anti-arsenic of 4-
N, the two ethyls of N--m-toluidine
The amino anti-arsenic of 4-
2, the two chlorine carbolic acids of 4-
The amino anti-arsenic of 4-
2,4,6-three bromo-3-hydroxyl-benzene sulfonic acid
The amino anti-arsenic of 4-
3, the two chlorine carbolic acid sulfonic acid of 5-
The amino anti-arsenic of 4-
3, the two chloro-2-hydroxyl-benzene sulfonic acids of 5-
The amino anti-arsenic of 4-
N-ethyl-N-(2-Hydroxyalkyl base-3-thiopropyl)-m-toluidine sodium salt
The amino anti-arsenic of 4-
Sa Xiu Hydroxyalkyl yl benzoic acid
The amino anti-arsenic of 4-
Two methylanilines
The amino anti-arsenic of 4-
N-ethyl-N-(2-Hydroxyalkyl base-3-thiopropyl)-m-toluidine
The amino anti-arsenic of 4-
2,2 '-AZINO-two (3-ethylbenzene thiazole-6-sulfonic acid)
The amino anti-arsenic of 4-
4-Hydroxyalkyl base-3-methoxy benzoic acid
The amino anti-arsenic of 4-
3-methyl-ethyl-Hydroxyalkyl base aniline.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102298026A (en) * | 2010-06-25 | 2011-12-28 | 苏州艾杰生物科技有限公司 | Method for determining glycine and glycine determination kit |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102298026A (en) * | 2010-06-25 | 2011-12-28 | 苏州艾杰生物科技有限公司 | Method for determining glycine and glycine determination kit |
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Open date: 20091223 |