31 results about "Creatininase" patented technology

The present invention generally relates to the treatment of uremic toxins in vivo using uremic toxin-treating enzymes, and/or cells capable of producing uremic toxin-treating enzymes or otherwise reacting with uremic toxins. Non-limiting examples of cases where the treatment of uremic toxins is desired include renal disease or dysfunction, gout, subjects receiving chemotherapy, or the like. In one aspect, the treatment includes an oral delivery composition able to reduce the blood concentration of one or more non-protein nitrogen compounds in vivo. The composition, in some cases, may comprise one, two, or more uremic toxin-treating enzymes, such as urease, uricase or creatininase. The oral delivery composition may be able to deliver the uremic toxin-treating enzymes, substantially undigested, to the intestines, where the enzymes can interact with uremic toxins transported to the intestines from the bloodstream. In another aspect, the treatment includes an oral delivery composition comprising a cell able to reduce the concentration of one or more uremic toxins in vivo. In some cases, the cell may be designed to overexpress one, two, or more uremic toxin-treating enzymes, such as urease, uricase or creatininase, for example, by transfecting the cell with a corresponding gene. In some embodiments, a species able to react with or otherwise sequester by-products of the uremic toxin-treating enzyme reactions may be included with the oral delivery composition. For example, if the by-product is ammonium, the species may be a sorbent able to adsorb ammonium, an enzyme able to react with the ammonium, or the like.

The invention discloses a detection kit for determining the content of creatinine in serum by an enzymic method. The detection kit comprises a reagent R1 and a reagent R2, wherein the reagent R1 contains 3-10g/L of buffering solution with the pH value being 7.5-8.0, 0.2-0.5g/L of EDTA, 0.5-2g/L of N-ethyl-N-(2-hydroxyl-3-sulfopropyl) m-toluidine sodium salt, 1 per mill-5 per mill of a surface active agent, 0.2-1g/L of sarcosine oxidase, 1-5KU/L of ascorbate oxidase, 2-5KU/L of creatinase and 0.5-1g/L of a stabilizing agent; the reagent R2 contains 3-10g/L of buffering solution with the pH value being 7.5-8.0, 0.1-0.5g/L of 4-aminoantipyrine, 0.1-0.4g/L of potassium ferricyanide, 2-8g/L of creatininase amidohydrolase, 1-6KU/L of peroxidase and 0.5-2g/L of a preservative. The detection kit disclosed by the invention is excellent in stability, strong in interference resistance and high in clinical application value.

The invention relates to a composition for eliminating the interference of a calcium dobesilate medicine on creatininase-method detection. The composition comprises an R1 reagent, an R2 reagent and ahigh molecular oxidant, wherein the high molecular oxidant is one or more of high molecular peroxy acid, high molecular selenium oxide, high molecular sulfur chloride ether, N-substituted imide, water-soluble tetrazole and a Dess-Martin oxidant. The high molecular oxidant in the composition disclosed by the invention can react with the calcium dobesilate in a sample to be detected, the calcium dobesilate loses the effect of interfering enzyme-method creatinine detection and the accuracy of sarcosine-oxidase-method detection is improved, so that the problem of serious negative interference, caused by high-concentration calcium dobesilate in blood of a patient, on an enzyme-method creatinine project detection result when the patient orally takes the calcium dobesilate medicine is solved.

The invention is about a method of measuring the content of creatinine, and it also concerns the reagent box of creatinine diagnosis. This invention belongs to the field of medical testing and measuring technology. The reagent box is consisted of buffer solution, creatinine enzyme, creatine enzyme, sarcosine onidase, anaerooxidase and stabilizer. Firstly, we cause an enzyme-coupled reaction through mixing the sample and the reagent according to a certain proportion of volume; secondly, put the final reactant under the biochemical analyzer and test the absorbance variational situation (speed) of dominant wavelength; then we can get the content of creatinine. By using this invention, we can get the necessary measuring result with high sensitiveness and fine precision through biochemical analyzer, and the result would not be contaminated by material of internal and exogenous sources. Thus, this method can be conveniently promoted and applied.

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.

The invention provides a creatinine detection kit and a use method thereof. The kit comprises a reagent R1 and a reagent R2, wherein the reagent R1 comprises creatinase, sarcosine oxidase, peroxidase,ascorbic acid oxidase, 2, 4, 6-tribromo-3-hydroxybenzoic acid and 3, 5-dichloro-2-hydroxybenzenesulfonic acid; and the reagent R2 comprises creatinase, peroxidase, potassium ferrocyanide and 4-aminoantipyrine. The creatinine detection kit provided by the invention can resist interference of etamsylate with the concentration of 300mg/L (the highest blood concentration) or below in a to-be-detectedsample, has a wider linear range (0-6000 millimoles/L), is applicable to detection of samples with high creatinine content and can correctly evaluate renal functions of surgical patients.

The invention relates to the technical field of biology, and particularly relates to a reagent combination, a reagent or a kit for measuring creatinine content. The reagent combination comprises a reagent group 1 and a reagent group 2; the reagent group 1 comprises creatinase, sarcosine oxidase, ascorbic acid oxidase, chromogen and catalase; the reagent group 2 comprises creatinase, 4-aminoantipyrine and peroxidase; and the chromogen is selected from N-(2-hydroxy-3-sulfopropyl)-3, 5-dimethoxyaniline or sodium salt thereof and N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3-5-dimethoxyaniline. An inhibitor is added into the measured reagent, so that generation of neocreatine and creatinine in the process of measuring is blocked, and interference existing in a sample in creatinine detection is lowered.

An Escherichia coli engineering bacterium for expressing Pseudomonas putida creatinase, the Escherichia coli engineering bacterium is obtained by transforming an expression vector containing the following foreign DNA into Escherichia coli: the foreign DNA is putida The DNA coding sequence of Pseudomonas putida creatinase, the amino acid sequence of Pseudomonas putida creatinase is shown in Seq No.2. The present invention also provides a construction method of the Escherichia coli engineering bacterium and the application of the recombinant Pseudomonas putida creatinase obtained by fermentation of the Escherichia coli engineering bacterium of the present invention in creatinine detection. The Escherichia coli engineering bacterium of the present invention is fast and simple to cultivate, the induction conditions are easy to control, the production cost is low, and it has the potential of industrial production; meanwhile, the recombinant Pseudomonas putida creatinase prepared by fermentation thereof has good creatinase activity and exhibits Excellent temperature stability, heat resistance and pH stability.

The invention discloses a creatinine detection reagent capable of resisting interference of medicines such as calcium dobesilate. The creatinine detection reagent comprises a first reagent and a second reagent, wherein the first reagent comprises 3-(N-morpholinyl)propanesulfonic acid, TOOS, gentamicin, sarcosine oxidase, creatinase, ascorbic acid oxidase and catalase, and the second reagent comprises 3-(N-morpholinyl)propanesulfonic acid, 4-AAP, creatininase amidohydrolase, peroxidase, preservative and potassium ferrocyanide; the first reagent and/or the second reagent further comprise/comprises piperidine oxygen radicals. According to the creatinine detection reagent provided by the invention, the interference effect of medicines such as calcium dobesilate can be reduced, the accuracy ofcreatinine detection in clinical detection is ensured, and a creatinine detection method with a wide linear range, strong medicine interference resistance and high accuracy is realized.

A yeast engineering bacterium for expressing Pseudomonas putida creatinase, which is obtained by transforming an expression vector containing the following foreign DNA into Pichia pastoris: the foreign DNA is Pseudomonas putida The DNA coding sequence of Pseudomonas putida creatinase, the amino acid sequence of Pseudomonas putida creatinase is shown in Seq No.2. The present invention also provides the construction method of the yeast engineering bacterium and the application of the recombinant Pseudomonas putida creatinase fermented by the yeast engineering bacterium of the present invention in creatinine detection. The yeast engineering bacterium of the present invention is quick and easy to cultivate, the induction conditions are easy to control, the production cost is low, and it has the potential of industrial production; meanwhile, the recombinant Pseudomonas putida creatinase prepared by fermentation thereof has good creatinase activity and exhibits excellent Excellent temperature stability, heat resistance and pH stability.

The invention provides an electrode for detecting creatinine by an electrochemical method. The electrode comprises an electrode material base layer, the outer surface of the electrode material base layer is covered with an electron mediator enhancement layer; an enzyme composition capable of reacting with creatinine to generate hydrogen peroxide is fixed on the outer surface of the electron mediator enhancement layer; the electrode material base layer is composed of carbon paste or graphite containing Prussian blue; the enzyme composition is prepared from creatininase, creatinase and sarcosine oxidase according to the ratio of (3.75-6.25) to (1.25-2) to (0.75-1.25). The invention also provides test paper and a biosensor based on the electrode. The invention also provides a preparation method for preparing the electrode and the test paper, and a method for detecting the creatinine level by using the biosensor. The test paper provided by the invention can be used in various portable and rapid small electrochemical detection devices, and can realize rapid and convenient clinical detection or home monitoring of blood and urine creatinine.

The invention discloses a serum creatinine detection card for optical differential signal processing and a preparation method and application thereof.The serum creatinine detection card is composed of an upper card shell, a lower card shell, a diffusion layer, a filtering reaction layer, a separation reaction layer and a chromogenic reaction layer, the filtering reaction layer is provided with creatininase and creatine hydrolase, the separation reaction layer is provided with sarcosine oxidase and anti-interference enzyme liquid, and the chromogenic reaction layer is provided with an anti-interference enzyme liquid. Chromatogen substrates and catalase are arranged on the chromogenic reaction layer, serum is obtained through treatment and separation of the diffusion layer, the filtering reaction layer and the separation reaction layer, creatininase method step-by-step reactions are carried out on different layers respectively, enzyme liquid is arranged in a layered mode, and the substrate concentration and the enzyme liquid concentration are beneficial to full reaction of all steps; reaction by-products of all the steps cannot influence enzyme liquid of other reaction steps and the reaction process, interference is reduced, creatinine and creatine are reacted and converted to the maximum extent, the content of serum creatinine is rapidly, accurately and quantitatively detected, the accuracy and stability of the detection result are greatly improved, use is easy, and operation by professionals is not needed.

The invention relates to a method for measuring the creatinine content and a creatinine diagnosis agent box in the field of medical testing technology. The agent box comprises: buffer solution, adenosine triphosphate, phosphoenolpyruvate, reducing coenzyme, atinase enzyme, creatine-kinase, pyruvate kinase, large document handler and stabilizer. It mixes the sample and the agent with a certain volume ratio to do enzyme coupling reaction; then it dispositions the end resting material on the chemical analyzer to detect the speed of the main wavelength light absorption to measure the content of the creatinine.

The invention discloses a preservative composition for the antisepsis of an enzyme creatinine agent, wherein the preservative composition comprises the following raw materials by weight: 1-3 parts ofa stabilizer, 0.4-1.3 parts of a preservative, 1-2 parts of a surfactant, 1-1.3 parts of a washing agent, 1-2 parts of ascorbate oxidase creatininase, 0.5-1.5 parts of sarcosine oxidase, 2-3 parts ofa buffer liquid, 2-3 parts of peroxidase, 3-5 parts of aminoantipyrine, and 2-4 parts of potassium ferrocyanide. The method specifically comprises: 1, respectively placing 3 parts of thiosulfate, 3 parts of thiocyanate, 3 parts of maleic acid, and 3 parts of sodium dialkyl sulfate in a mixing device, and uniformly stirring by a stirring rod. The invention relates to the technical field of creatinine. According to the present invention, with the preservative composition, the enzyme creatinine reagent can be prevented from being corroded by other corrosive chemical substances during the quantitative determination of the creatinine in human serum, plasma (lithium heparin and K2EDTA) and urine by the ADVIA biochemical analysis system so as not to affect the determination results.