CN102375066A - Creatinine content detecting reagent and kit, and manufacturing and using methods of kit - Google Patents
Creatinine content detecting reagent and kit, and manufacturing and using methods of kit Download PDFInfo
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- CN102375066A CN102375066A CN2011102799693A CN201110279969A CN102375066A CN 102375066 A CN102375066 A CN 102375066A CN 2011102799693 A CN2011102799693 A CN 2011102799693A CN 201110279969 A CN201110279969 A CN 201110279969A CN 102375066 A CN102375066 A CN 102375066A
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Abstract
The invention relates to a creatinine content detecting reagent and kit and manufacturing and using methods of the kit. The creatinine content detecting reagent comprises creatininase, creatinase, creatine oxidase and horseradish peroxidase; and a fluorescent dye, i.e., 5(6)-carboxyl-2,7-dichlordihydro fluorescein ester dipivalate comprises a dye bottom layer, a protease fixing layer and a top layer protecting layer. The using method has high selectivity; a protease has high specificity to reactants; the creatininase can be only used for catalyzing the hydrolysis reaction of creatinine, and does not have catalysis activity on other reaction substrates; three enzymes with high specificity are related to the detection process, so that the selectivity of the method is increased exponentially; the response fed by the kit is fluorescent response; and compared with certain methods for detecting color variations, the invention has the advantages of high sensitivity and interference resistance under the action of the inherent characteristics of fluorescent response.
Description
Technical field
The invention belongs to the kreatinin detection range, especially a kind of kreatinin content detecting reagent and kit, and the making and use method of kit.
Background technology
Kreatinin is the product of phosphocreatine metabolism, and its chemical constitution is that creatine passes through the lactams that dehydrating condensation forms.Kreatinin generally is present in the blood, arrives kidney and is filtered through blood circulation.For the normal crowd of renal function, the kreatinin concentration in the blood varies in different localities because of the people.Generally speaking, under the situation of no big physical load, the kreatinin concentration in women's the blood is at 45-60 μ M, and the male sex's blood kreatinin concentration is floated in the scope of 60-110 μ M.When renal function goes wrong; Kidney filters the ability of removing kreatinin will be reduced, and causes the concentration abnormality of kreatinin in the blood to raise and kreatinin concentration reduction in the urine, therefore through measuring the concentration of kreatinin in blood and the urine; Just can measure creatinine clearance even glomerulus rate filters; These two parameters have great importance for weighing renal function, therefore, can measure the kreatinin concentration in blood and the urine efficiently; For clinical and scientific research, all has very great application prospect.
The measurement of kreatinin is difficulty relatively.Method commonly used at present is mass spectroscopy (isotope dilution mass spectrometry), high performance liquid chromatography, liquid chromatography mass logotype method and antibody test (ELISA); These methods generally all need expensive instrument; Like mass spectrometer, liquid chromatograph or ELIASA etc.; And the operation that received the analytical chemistry operating personnel of systematic training, greatly improve financial cost and time cost that kreatinin is measured, and kreatinin is measured in backwoodsman application.
Mass spectroscopy is through containing the kreatinin of certain isotope (like 0-18) and this sample is carried out mass spectrophotometry artificial adding the in sample.The abundance of 0-18 and 0-18 can calculate kreatinin in the abundance of occurring in nature concentration among the comparison mass spectrum result.The artificial kreatinin that adds can strict be controlled, and as a kind of internal standard (internal calibration), this method can effectively reduce error.According to current research, on the low side with the kreatinin concentration that mass spectroscopy obtains, this will cause a series of problems clinically, and U.S. FDA also begins to consider new kreatinin detection means.
Liquid phase chromatography is that the use high performance liquid chromatography carries out purifying to sample and detects with mass spectrum on mass spectral basis.Mass spectroscopy and liquid phase chromatography can't robotizations, complicated operation; And need to analyze team instrument is carried out operation and maintenance; And the ELISA method need be used expensive ELIASA, though its sensitivity is high, selectivity is relatively poor; Over-evaluate the concentration of kreatinin through regular meeting, cause the excessive or not enough of clinical application.
Summary of the invention
The objective of the invention is to overcome the weak point of prior art, provide a kind of selectivity kreatinin content detecting reagent and kit high, highly sensitive and easy to use, and the making and use method of kit.
The objective of the invention is to realize through following technical scheme:
A kind of kreatinin content detecting reagent; Comprise creatininase, kreatinase, sarcosine oxidase and horseradish peroxidase; Fluorescent dye 5 (6)-carboxyls-2,7-dichloro-dihydro luciferin two pivalates, kreatinin are hydrolyzed under the catalysis of creatininase and are creatine; Creatine is hydrolyzed into methyl amimoacetic acid under the catalysis of kreatinase; Methyl amimoacetic acid generates glycocoll and a spot of hydrogen peroxide thereupon under the catalysis of sarcosine oxidase, under the catalysis of horseradish oxide enzyme, catalytic oxidation of hydrogen peroxide 5 (6)-carboxyls-2; 7-dichloro-dihydro luciferin two pivalates also generate the product with fluorescence response; Speed and typical curve through monitoring fluorescence strengthens obtain kreatinin concentration, and wherein said creatininase: kreatinase: sarcosine oxidase: horseradish peroxidase: the weight ratio of fluorescent dye is: 1: 50: 60: 5.
And, comprise following three-decker:
(1) dyestuff bottom: cellulose and mixing of D4 hydrogel and stirring with the fluorescent dye modified are coated with the film of last layer D4 hydrogel and make it natural air drying with the scraper film applicator on a clean plastic sheeting;
(2) proteinase fixed bed: in the WS of PVA-Sbq (POLYPROPYLENE GLYCOL phenylpyridine ethene complex compound), add 10% bovine serum albumin and react required creatininase, kreatinase, sarcosine oxidase and horseradish peroxidase; Stir more than 25-40 minute; Be coated with the PVA-Sbq solution of last layer enzyme with the scraper film applicator; Uviol lamp is placed the top 30 minutes of rete, make phenylpyridine ethene crosslinked and rete is become dry
(3) top layer protective seam: in the above with scraper film applicator be coated with D6 hydrogel that last layer contain 10% carbon black at last, and make the rete natural air drying.
And the concrete preparation process of three-decker is following:
(1) 5 (6)-carboxyls-2 of the 176mg cellulose being fixed, the 4%D4 hydrogel solution mixing of 7-dichloro-dihydro luciferin two pivalates and 2.6g also stirs, and cellulose powder is evenly disperseed in solution,
(2) evenly on plastic sheeting, to make it to form a layer thickness be the film of 100 μ M and dry with prepared solution in the step (1);
(3) with 30mg kreatinase, 35mg horseradish peroxidase, 0.63mg creatininase, 2.6mg sarcosine oxidase, 340mg water and 1.33g concentration be 10% and the PVA-Sbq solution that contains 10% bovine serum albumin mix, and stir on ice:
(4) in step (2), coat PVA-Sbq/ enzyme solutions prepared in the step (3) on the film of preparation, the thickness of rete is 100 μ m, and film was placed uviol lamp following 30 minutes;
(5) in step (4), be coated with solution and the natural air drying that last layer contains 6% carbon black and 1.8%D6 on the prepared film.
A kind of kreatinin reagent box for detecting content comprises above-mentioned detectable.
A kind of preparation method of kreatinin reagent box for detecting content is characterized in that: step is following:
(1) 5 (6)-carboxyls-2 of the 176mg cellulose being fixed, the 4%D4 hydrogel solution mixing of 7-dichloro-dihydro luciferin two pivalates and 2.6g also stirs, and cellulose powder is evenly disperseed in solution;
(2) evenly on plastic sheeting, to make it to form a layer thickness be the film of 100 μ M and dry with prepared solution in the step (1);
(3) with 30mg kreatinase, 35mg horseradish peroxidase, 0.63mg creatininase, 2.6mg sarcosine oxidase, 340mg water and 1.33g concentration be 10% and the PVA-Sbq solution that contains 10% bovine serum albumin mix, and stir on ice;
(4) in step (2), coat PVA-Sbq/ enzyme solutions prepared in the step (3) on the film of preparation, the thickness of rete is 100 μ m, and film was placed uviol lamp following 30 minutes;
(5) in step (4), be coated with solution and the natural air drying that last layer contains 6% carbon black and 1.8%D6 on the prepared film.
A kind of method of application of kit; It is characterized in that: containing 100mMHEPES, 150mM sodium chloride, 5mM potassium chloride; 1.3mM in the damping fluid of lime chloride; PH7.3 adds the variable concentrations kreatinin, drives the sample solution of 150 μ L and assign on the kit and with luminoscope with kapillary two to begin to measure.
Advantage of the present invention and beneficial effect are:
1, method used in the present invention has the selectivity of height; As the protein enzyme, it has the specificity of height to reactant, the hydrolysis reaction that creatininase can only the catalysis kreatinin; Reaction substrate to other does not all have catalytic activity; Owing to relate to the enzyme of three kinds of high degree of specificity in the testing process, the selectivity of this method is index especially to be increased, and the response that kit provides at last is a fluorescence response; Compare with some method that detects change color, the inherent characteristic of fluorescence response is given higher sensitivity of the present invention and anti-jamming capacity.
2, the application provides a kind of novel being used for to detect the kit of blood and urine kreatinin content; This kit will have specific enzyme and the superoxide fluorescent dye sensitive will be fixed on the kit kreatinin; And enzymatic reaction conversion is fluorescence signal and is able to detect; This kit can detect the above kreatinin of 10 μ M, meets request for utilization clinically.
3, enzyme used in the present invention has four kinds: creatininase, kreatinase, sarcosine oxidase and horseradish peroxidase; Kreatinin in the sample is degraded to glycocoll through the first three enzyme, generates a spot of hydrogen peroxide simultaneously, under the catalysis of horseradish peroxidase; Hydrogen peroxide can a kind of script of oxidation have the molecule of fluorescence; Reaction product is the derivant of luciferin, has stronger fluorescence response, through detecting the enhancing speed of fluorescence; Can detect the generating rate of hydrogen peroxide, and indirect calculation goes out the concentration of kreatinin.
4, the present invention compares the kit that existing kreatinin detection technique and the application are set forth; Important parameter as renal function; The creatine that creatinine clearance and glomerulus rate are filtered need repeatedly carry out the kreatinin of blood and urine and measure; Therefore, simple, the cheap and characteristics that have a high selectivity of the use that had of the mentioned kreatinin kit of the application will have very significant meaning.
The comparison of the existing kreatinin detection method of table 1
Description of drawings
Fig. 1 is the cross-linking reaction formula of phenyl vinylpyridine of the present invention;
Fig. 2 is the structure of the kit of creatinine sensor use of the present invention;
Fig. 3 is the fluorescence response in time of the present invention's sample of kit under different potassium concentrations; Fig. 3-1 and Fig. 3-2 are respectively corresponding with the dual mode express time.
Fig. 4 gain in strength for the fluorescence that calculates according to Fig. 4 and potassium concentration between correlativity.
Embodiment
Below in conjunction with embodiment, the present invention is further specified, following embodiment is illustrative, is not determinate, can not limit protection scope of the present invention with following embodiment.
If the number percent that following examples are mentioned does not have special indicating all to be weight percentage, the mM of unit representes every liter of mM.
One, the principles of chemistry of kreatinin kit
The kreatinin kit that the present invention set forth comprises four kinds of enzymes and a kind of fluorescent dye, and they are respectively: creatininase, kreatinase, sarcosine oxidase and horseradish peroxidase.The chemical name of fluorescent dye is 5 (6)-carboxyls-2,7-dichloro-dihydro luciferin two pivalates (the following A of structure), and the fluorescent yield of this fluorescent dye is very low; Therefore under general condition; Its fluorescent emission efficient is very low, and this fluorescent dye can be oxidized into 2,7-dichlorofluorescein (structural formula is seen B); 2; The 7-dichlorofluorescein has very high quantum yield, so these fluorescent dye 5 (6)-carboxyls-2, and the oxidizing process of 7-dichloro-dihydro luciferin two pivalates can be monitored through the mensuration to fluorescence.
5 (6)-carboxyls-2, the structure of 7-chlorine dihydrofluorescein two pivalates (A) and oxidation afterproduct (B) thereof
The present invention narrates as follows the chemical process of the detection of kreatinin: kreatinin is hydrolyzed under the catalysis of creatininase and is creatine; Creatine is hydrolyzed into methyl amimoacetic acid under the catalysis of kreatinase; Methyl amimoacetic acid generates glycocoll and a spot of hydrogen peroxide (reaction equation of above process is following) thereupon under the catalysis of sarcosine oxidase; Under the catalysis of horseradish oxide enzyme, the catalytic oxidation of hydrogen peroxide that forms in the said process 5 (6)-carboxyls-2,7-dichloro-dihydro luciferin two pivalates also generate the product with fluorescence response; Through speed and the typical curve that monitoring fluorescence strengthens, the information of acquisition kreatinin concentration that can be indirect.
Two, protein enzyme and fluorescent dye is fixing
In kit, enzyme and fluorescent dye will be fixed in the solid phase substrate.The bottom is that (a kind of polyurethane polyureas monoethylene glycol hydrogel, specifically see following patent: supporting layer US 2008/0152864A1), principal ingredient are the linear polymers of polyglycol and polyesteramide to the D4 hydrogel.In view of the instability of fluorescent dye (structural formula is A), it is fixed on the cellulose through covalent bond, and the cellulosic particle of fluorescent dye is dispersed in the D4 hydrogel layer uniformly.The protein enzyme is fixed on the second layer, and its principal ingredient is the multipolymer of polyvinyl alcohol (PVA) and phenyl vinylpyridine.The hydrosol of polyvinyl alcohol (PVA) phenyl vinylpyridine with become collosol state after the protein enzyme mixes.After potpourri was by ultraviolet irradiation, the phenyl vinylpyridine produced crosslinked, makes mixture solidified; And the protein enzyme also will be by physical fixation in gel, and its reaction equation is seen Fig. 1.
Owing to the protein enzyme is fixed in the solid and the preservation that is dried, so its hydrolysis comparatively speaking can be difficult a lot of with degraded: at first owing to lack of water, the possibility that hydrolysis takes place reduces greatly; Secondly owing to the existence of solid phase protective seam, protein decomposition enzyme contacted protein molecular proportion is difficulty, and the hydrolysis reaction that results in its catalysis can not take place, and therefore can expect that the protein enzyme that is fixed is also with more stable.
Three, the method for making of kit:
(1) dyestuff bottom: the cellulose of fluorescent dye modified and D4 hydrogel are mixed and stirred a hour, on a clean plastic sheeting, be coated with the film of last layer D4 hydrogel and make it natural air drying immediately with the scraper film applicator.
(2) proteinase fixed bed: in the WS of PVA-Sbq (POLYPROPYLENE GLYCOL phenylpyridine ethene complex compound), sneak into 10% bovine serum albumin and react required four kinds of enzymes (creatininase, kreatinase, sarcosine oxidase and horseradish peroxidase) and stir more than 30 minutes; This solution will be used for the immobilization of enzyme; Be coated with the PVA-Sbq solution of last layer enzyme with the scraper film applicator; Uviol lamp is placed the top 30 minutes of rete, make phenylpyridine ethene crosslinked and rete is become dry.
US 2008/0152864A1) and make the rete natural air drying (3) top layer protective seam: last be coated with the D6 hydrogel that last layer contains 10% carbon black with the scraper film applicator in the above (a kind of polyurethane-polyglycol hydrogel is specifically seen following patent:.
(4) using cutting machine to extract diameter is about the disk of 5mm and is installed in the sky kit.The shape of last kit is seen Fig. 2.Blood sample gets into kit and the valve through black through kapillary from the kit right part and contacts with sensor and produce signal and received by luminoscope.The position of having reserved other five other sensors at present on the kit has reached the purpose of a plurality of detection things of one-time detection.
Four, specifically prepare instance:
1, kit preparation process:
(1) 5 (6)-carboxyls-2 of the 176mg cellulose being fixed, the 4%D4 hydrogel solution mixing of 7-dichloro-dihydro luciferin two pivalates and 2.6g also stirs, and cellulose powder is evenly disperseed in solution,
(2) evenly on plastic sheeting, to make it to form a layer thickness be the film of 100 μ M and dry with prepared solution in the step (1);
(3) with 30mg kreatinase, 35mg horseradish peroxidase, 0.63mg creatininase, 2.6mg sarcosine oxidase, 340mg water and 1.33g concentration be 10% and the PVA-Sbq solution that contains 10% bovine serum albumin mix, and stir on ice:
(4) in step (2), coat PVA-Sbq/ enzyme solutions prepared in the step (3) on the film of preparation, the thickness of rete is 100 μ m, and film was placed uviol lamp following 30 minutes;
(5) in step (4), be coated with solution and the natural air drying that 6% carbon black and 1.8%D6 are contained in the upper strata on the prepared film;
(6) extract in the step (5) prepared film and be installed on the kit and can measure.
In view of the instability of fluorescent dye (structural formula is A), it is fixed on the cellulose through covalent bond, and fixing means is following:
1g aminocellulose and 50mg fluorescent dye are mixed in solvent dimethylformamide and stir; The dicyclohexylcarbodiimide and the 100mg N-hydroxy-succinamide that slowly add about 200mg, filtration and also dry after stirring the mixture four hours with dimethyl imide flushing filter residue.
2, sample measurement:
Containing 100mMHEPES; 150mM sodium chloride; 5mM potassium chloride, (pH7.3) adds the variable concentrations kreatinin in the damping fluid of 1.3mM lime chloride, drives the sample solution of about 150 μ L and assign on the kit and with portable luminoscope OPTTCCA-TS with kapillary two to begin to measure.
3, data analysis:
The computer that links to each other with luminoscope will write down the photo emissions intensity of reaction generation and the curve of time, can be according to this curve in the hope of reaction rate, and this speed is exactly the response of kit to kreatinin, can be in the hope of the concentration of kreatinin according to typical curve.
4, experimental data: kit is to the response of kreatinin:
Response shows at Fig. 3, Fig. 4 the kit that above-mentioned steps obtains to kreatinin.Fig. 3 is the fluorescence response in time of the sample of kit under different kreatinin concentration.Can see that under different kreatinin concentration, the speed that fluorescence rises is different.When not having kreatinin in the solution, fluorescence intensity has risen about 40% in 60 seconds.And when containing the kreatinin of 0.5mM in the solution, fluorescence intensity has risen about 80% in 60 seconds.So the speed that fluorescence strengthens and the concentration of kreatinin are closely-related, the speed that also can use fluorescence to increase is calculated the concentration of kreatinin.
Fig. 4 represent according to the fluorescence that Fig. 3 calculates gain in strength and kreatinin concentration between correlativity.Can more significantly see the correlativity that kreatinin and fluorescence are advanced the speed.Kreatinin concentration and fluorescence are advanced the speed linear.This will greatly facilitate the calculating of typical curve.The sensitivity of calculating gained according to 3 σ methods is 10 μ M.
5, conclusion
Present patent application has been set forth and has been planted the kit that novel being used for detects blood and urine kreatinin content.This kit will have specific enzyme and the superoxide fluorescent dye sensitive will be fixed on the kit kreatinin, and enzymatic reaction conversion is fluorescence signal and is able to detect.
This kit can detect the above kreatinin of 10 μ M at an easy rate, meets request for utilization clinically.
Claims (6)
1. kreatinin content detecting reagent; It is characterized in that: comprise creatininase, kreatinase, sarcosine oxidase and horseradish peroxidase; Fluorescent dye 5 (6)-carboxyls-2,7-dichloro-dihydro luciferin two pivalates, kreatinin are hydrolyzed under the catalysis of creatininase and are creatine; Creatine is hydrolyzed into methyl amimoacetic acid under the catalysis of kreatinase; Methyl amimoacetic acid generates glycocoll and a spot of hydrogen peroxide thereupon under the catalysis of sarcosine oxidase, under the catalysis of horseradish oxide enzyme, catalytic oxidation of hydrogen peroxide 5 (6)-carboxyls-2; 7-dichloro-dihydro luciferin two pivalates also generate the product with fluorescence response; Speed and typical curve through monitoring fluorescence strengthens obtain kreatinin concentration, and wherein said creatininase: kreatinase: sarcosine oxidase: horseradish peroxidase: the weight ratio of fluorescent dye is: 1: 50: 60: 5.
2. kreatinin content detecting reagent according to claim 1 is characterized in that: comprise following three-decker:
(1) dyestuff bottom: cellulose and mixing of D4 hydrogel and stirring with the fluorescent dye modified are coated with the film of last layer D4 hydrogel and make it natural air drying with the scraper film applicator on a clean plastic sheeting;
(2) proteinase fixed bed: in the WS of PVA-Sbq (POLYPROPYLENE GLYCOL-phenylpyridine ethene complex compound), add 10% bovine serum albumin and react required creatininase, kreatinase, sarcosine oxidase and horseradish peroxidase; Stir more than 25-40 minute; Be coated with the PVA-Sbq solution of last layer enzyme with the scraper film applicator; Uviol lamp is placed the top 30 minutes of rete, make phenylpyridine ethene crosslinked and rete is become dry;
(3) top layer protective seam: in the above with scraper film applicator be coated with D6 hydrogel that last layer contain 10% carbon black at last, and make the rete natural air drying.
3. kreatinin content detecting reagent according to claim 2 is characterized in that: the concrete preparation process of three-decker is following:
(1) 5 (6)-carboxyls-2 of the 176mg cellulose being fixed, the 4%D4 hydrogel solution mixing of 7-dichloro-dihydro luciferin two pivalates and 2.6g also stirs, and cellulose powder is evenly disperseed in solution;
(2) evenly on plastic sheeting, to make it to form a layer thickness be the film of 100uM and dry with prepared solution in the step (1);
(3) with 30mg kreatinase, 35mg horseradish peroxidase, 0.63mg creatininase, 2.6mg sarcosine oxidase, 340mg water and 1.33g concentration be 10% and the PVA-Sbq solution that contains 10% bovine serum albumin mix, and stir on ice;
(4) in step (2), coat PVA-Sbq/ enzyme solutions prepared in the step (3) on the film of preparation, the thickness of rete is 100 μ m, and film was placed uviol lamp following 30 minutes;
(5) in step (4), be coated with solution and the natural air drying that last layer contains 6% carbon black and 1.8%D6 on the prepared film.
4. a kreatinin reagent box for detecting content is characterized in that: comprise claim 1 or 2 or 3 described detectable.
5. the preparation method of a kreatinin reagent box for detecting content as claimed in claim 4, it is characterized in that: step is following:
(1) 5 (6)-carboxyls-2 of the 176mg cellulose being fixed, the 4%D4 hydrogel solution mixing of 7-dichloro-dihydro luciferin two pivalates and 2.6g also stirs, and cellulose powder is evenly disperseed in solution;
(2) evenly on plastic sheeting, to make it to form a layer thickness be the film of 100uM and dry with prepared solution in the step (1);
(3) with 30mg kreatinase, 35mg horseradish peroxidase, 0.63mg creatininase, 2.6mg sarcosine oxidase, 340mg water and 1.33g concentration be 10% and the PVA-Sbq solution that contains 10% bovine serum albumin mix, and stir on ice;
(4) in step (2), coat PVA-Sbq/ enzyme solutions prepared in the step (3) on the film of preparation, the thickness of rete is 100 μ m, and film was placed uviol lamp following 30 minutes;
(5) in step (4), be coated with solution and the natural air drying that last layer contains 6% carbon black and 1.8%D6 on the prepared film.
6. the method for application of a kit as claimed in claim 4; It is characterized in that: containing 100mM HEPES, 150mM sodium chloride, 5mM potassium chloride; 1.3mM in the damping fluid of lime chloride; PH 7.3, add the variable concentrations kreatinin, drive the sample solution of 150 μ L and assign on the kit and with luminoscope with kapillary two to begin to measure.
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| CN201310360937.5A CN103472041B (en) | 2011-09-20 | 2011-09-20 | A kind of using method of creatinine content detection kit |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103278468A (en) * | 2013-05-24 | 2013-09-04 | 宁波美康生物科技股份有限公司 | Creatinine detection reagent |
| CN107257855A (en) * | 2015-02-27 | 2017-10-17 | 雷迪奥米特医学公司 | Modified kreatinase |
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| CN104459164B (en) * | 2014-11-28 | 2016-03-23 | 山东博科生物产业有限公司 | A kind of serum creatinine detection reagent |
| CN106645758B (en) * | 2017-01-03 | 2019-03-22 | 长沙中生众捷生物技术有限公司 | The Test paper of creatinine |
| CN107760759A (en) * | 2017-11-07 | 2018-03-06 | 上海纳米技术及应用国家工程研究中心有限公司 | The method for detecting prostate cancer target methyl amimoacetic acid |
| CN114774256B (en) * | 2022-04-14 | 2023-03-24 | 重庆云芯医联科技有限公司 | Optical differential signal processing blood creatinine detection card, preparation method and application |
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| CN103278468A (en) * | 2013-05-24 | 2013-09-04 | 宁波美康生物科技股份有限公司 | Creatinine detection reagent |
| CN103278468B (en) * | 2013-05-24 | 2015-09-02 | 宁波美康生物科技股份有限公司 | A kind of creatinine detection reagent |
| CN107257855A (en) * | 2015-02-27 | 2017-10-17 | 雷迪奥米特医学公司 | Modified kreatinase |
| CN107257855B (en) * | 2015-02-27 | 2022-04-29 | 雷迪奥米特医学公司 | Modified creatinase |
| CN114958814A (en) * | 2015-02-27 | 2022-08-30 | 雷迪奥米特医学公司 | Modified creatinase |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103472239B (en) | 2016-03-09 |
| CN103472239A (en) | 2013-12-25 |
| CN102375066B (en) | 2013-10-16 |
| CN103472041B (en) | 2016-01-20 |
| CN103472041A (en) | 2013-12-25 |
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