145 results about "Glycated Hb" patented technology

A non-enzymatic electrochemical method of simultaneous measurement of hemoglobin (Hb) and percentage of glycated hemoglobin (% HbA1c) in a blood sample is disclosed. The method includes determining the total amount of hemoglobin in a sample by electrochemically measuring the voltammetric current due to iron (II) and iron (III) redox portions in hemoglobin and determining the percentage of glycated hemoglobin (HbA1c) by potentiometry. Also disclosed is a novel screen-printed electrode (SPE) strip modified for potentiometric measurement of HbA1c.

The invention provides enzymatic methods for direct determination of percentage of glycated hemoglobin in blood samples without the need of a separated measurement of total hemoglobin content in blood samples. The methods utilizes one or two different types of oxidizing agents which selectively oxidize low-molecular weight reducing substances and high-molecular weight (mainly hemoglobin) reducing substances in blood samples, coupled with enzymatic reactions catalyzed by proteases, fructosyl amino acid oxidase, and peroxidase. The invention provides kits for performing the methods of the invention.

The present invention provides an electrochemical detection method for content of glycosylated hemoglobin in the blood, and is characterized in that: whole blood specimen is diluted by combined liquid ranging from 30 times to 70 times and passes through a glycosylated protein recognizer, electric activated small molecules are filled into the effluent liquid from the glycosylated protein recognizer, and electrochemical response of the liquid is detected by chemically modified electrodes after the filling, thereby obtaining the concentration of the blood glucose. The glycosylated protein recognizer is eluted by elute solvent, and the electrochemical response of the elute solvent is detected and the concentration of the glycosylated hemoglobin is obtained, thereby obtaining the content of the glycosylated hemoglobin of the sample after calculation. As the electrochemical detection is simple in operation, lower in cost and rapid in the analysis, the present invention is expected to realize the rapid determination of the clinical glycosylated hemoglobin and the rapid determination of the glycosylated hemoglobin after separation and elution.

The invention relates to an enzymatic test kit for glycosylated hemoglobin, which is composed of hemolysis buffer solution, reagent R1a, reagent R1b and reagent 2; wherein, the hemolysis buffer solution is 10 to 1000mmol / L N- cycloethyl-2-aminoethanesulfonic acid, 10 to 1000mmol / L MOPS and 1 to 100g / L polyethylene glycol oxide dodecyl ether; the reagent R1a is 100 to 10000 kU / L metalloproteinase, 0.1 to 10mmol / L 2-(iodophenyl)-3-(2, 4- dinitrobenzene)-5-(2, 4-disulfophenyl)-2H tetrazole sodium salt; the reagent R1b is 0.1 to 10mmol / L MES and 0.5 to 50mmol / L CaCl2; the reagent 2 is 1 to 100kU / L fructose valine oxidase with high activity, 30 to 600kU / L POD, 0.01 to 0.5mmol / L DA-64 and 10 to 1000mmol / L Tris-HCl. The test kit coincides with the characteristics of rapidness, large batch size and economy which are required by the glycosylated hemoglobin clinical tests.

The invention discloses a method for measuring glycosylated hemoglobin and a kit. The kit comprises a kit body, reagent bottles, a bottle mat, a hinge and a kit cap, wherein the kit body is connected with the kit cap through the hinge; the four reagent bottles are respectively used for placing a magnetic particle suspension, an erythrocyte splitting liquor, a flushing liquid and signals. According to the invention, magnetic particles capable of quantitatively capturing glycosylated hemoglobin and non-glycosylated hemoglobin in a sample are adopted, and percentage of the glycosylated hemoglobin in the sample to the total hemoglobin can be calculated only by detecting the content of glycosylated hemoglobin on the magnetic particles and comparing the content with the standard work curve. According to the invention, the operation is simple, and the quantitative detection on the percentage composition of the glycosylated hemoglobin can be completed only with one operation; and biological raw materials such as antibodies are not needed, so that the price is low, and detecting instrument miniaturization is easy to realize.

The invention discloses recombinant human fibroblast growth factor 21 fusion protein and application thereof in preparation of medicine for treating metabolic diseases. A nucleotide sequence of coding genes of the recombinant human fibroblast growth factor 21 fusion protein and an expression vector are connected to obtain a recombinant expression vector; the recombinant expression vector converts host cells, high-expression positive host cells are screened out, the cells are cultured, the recombinant human fibroblast growth factor 21 fusion protein is induced to be expressed, thalli are collected, breaking, centrifuging, clarifying and purifying are carried out, and the target product recombinant human fibroblast growth factor 21 fusion protein is obtained. The invention further discloses the application of the recombinant human fibroblast growth factor 21 fusion protein in preparation of the medicine for treating metabolic diseases. The recombinant human fibroblast growth factor 21 fusion protein can well control blood sugar fluctuation and remarkably improve the level of glycosylated hemoglobin and triglyceride.

Described herein are systems and methods for assaying a sample to quantitatively determine the percentage of glycated hemoglobin in the sample.

There is provided a method for specifically determining a glycated β-chain N-terminal of glycated hemoglobin using enzymes without a separation operation, and a determination reagent kit therefor. A protease that cleaves a glycated amino acid and / or a glycated peptide from a glycated β-chain N-terminal without substantially cleaving a glycated amino acid or a glycated peptide from a glycated α-chain N-terminal of glycated hemoglobin or a fragment thereof is screened. The method of specifically determining a glycated β-chain N-terminal of glycated hemoglobin and the determination reagent kit are provided by using the protease obtained by the screening method. According to the present invention, a glycated β-chain N-terminal of glycated hemoglobin can specifically be determined without a separation operation.

The invention discloses an aptamer of glycosylated hemoglobin and a preparation method of the aptamer. The nucleotide sequence of the aptamer of the glycosylated hemoglobin GHb comprises any one of sequences from SEQ ID NO: 1-SEQ ID NO: 4. The preparation method of the aptamer of the glycosylated hemoglobin comprises the following steps of: building a random sequence oligonucleotide library, preparing a glycosylated hemoglobin-micro magnetic bead compound, initially screening target material-nucleic acid compound, preparing a secondary single stranded DNA (deoxyribonucleic acid) oligonucleotide library, screening, and identifying the aptamer of the glycosylated hemoglobin. The prepared aptamer is free from toxicity, small in molecular weight, and easy to synthesize and mark, only can the glycosylated hemoglobin GHb be specifically identified, and the other homologous non-glycosylated hemoglobin Hb can not be identified and combined, so that the aptamer can be taken as a molecular beacon for detecting the content of the glycosylated hemoglobin GHb.

This invention provides an amadoriase having high reactivity with α-fructosyl hexapeptide (αF6P) derived from the β-chain amino terminus of glycated hemoglobin (HbAlc), which enables satisfactory quantification of αF6P cleaved from HbAlc. A novel amadoriase is derived from the amadoriase of the genus Coniochaeta by substitution of one or more amino acids at positions selected from the group consisting of positions 62, 63, 102, 106, 110, 113, 355, and 419. Such amadoriase enables rapid, simple, accurate, and satisfactory quantification of αF6P cleaved from HbAlc. With the use of such amadoriase, a method for measurement of HbAlc by an enzymatic method and a kit for measurement of HbAlc that are based on the same principle as used in the conventional standard method and excellent in terms of the relevance with the standard method.

The invention provides enzymatic methods for direct determination of percentage of glycated hemoglobin in blood samples without the need of a separated measurement of total hemoglobin content in blood samples. The methods utilizes one or two different types of oxidizing agents which selectively oxidize low-molecular weight reducing substances and high-molecular weight (mainly hemoglobin) reducing substances in blood samples, coupled with enzymatic reactions catalyzed by proteases, fructosyl amino acid oxidase. The amount of hydrogen peroxide generated in the reaction is measured for determination of percentage of glycated hemoglobin in blood samples. The invention provides kits for performing the methods of the invention.

The invention relates to a method for quantitative measurement of glycated hemoglobin (HbAIc) in a blood sample containing hemoglobin molecules. In this method, the hemoglobin molecules are extracted from the blood sample, adsorbed onto a roughened metal surface and subjected to a Surface Enhanced Raman Scattering (SERS) measurement. The invention provides a universal method for the measurement of the HbAIc concentration in blood.

The invention provides an elution method for detection of glycosylated hemoglobin with liquid chramatography and inspection equipment thereof, the method including following steps: step (1): preparing eluents of three kinds of ion concentration; step (2): performing elution on ion exchange resin which contains a to-be-tested sample with the said eluents of three kinds of ion concentration; step (3): performing chromatographic analysis on the said liquid that undergoes elution, and determining the content of the elution component; step (4): determining the concentration of the glycosylated hemoglobin HbA1c. The method is capable of solving the problem of cross contamination caused in the detection of eluents of two kinds of ion concentration. Compared with the method using eluents of four kinds of ion concentration, the method simplifies the inspection procedure and the structure of the inspection equipment, improves the reliability of the operation of the equipment, reduces the cost of the equipment, and greatly improves the accuracy and stability of the inspection.

Disclosed is a hemolysis reagent composition for the quantitative analysis of glycated hemoglobin (HbAlc) via an enzymatic method. The hemolysis reagent compositions of the present invention containing a zwitterionic surfactant and a nitrous compound considerably accelerate hemolysis rate due to the presence of the zwitterionic surfactant, and also induce the N-terminal β-chain of hemoglobin released by hemolysis to be exposed to the outside of the hemoglobin molecule and assists the participation of a greater number of molecules in the reaction, thereby improving reaction sensitivity and reaction rate of the glycated hemoglobin, and the nitrous compound makes the protein structure of the hemoglobin flexibly modified so that the amino acid sequence of the N-terminal β-chain of hemoglobin can be readily cut off, thereby significantly reducing the entire time required for the measurement and improving the accuracy of the measurement.

The invention relates to a glycosylated hemoglobin immunochromatographic test strip and a preparation method thereof and a kit. The test strip comprises a substrate, and a sampling pad, a coating filmand an absorption pad which are arranged on the substrate, wherein the sampling pad, the coating film and the absorption pad are sequentially connected from one end of the substrate to the other endof the substrate; a marking area, a detection area and a quality control area are sequentially arranged on the coating film; the marking area is coated with a coupling compound marked by fluorescent latex particles, and the coupling compound is formed by coupling a first dog and cat HbA1c monoclonal antibody and a first complex; the detection area is coated with a second dog and cat HbA1c monoclonal antibody; the quality control area is coated with a second complex, and the first complex and the second complex can be specifically combined; and the first dog and cat HbA1c monoclonal antibody can be specifically combined with hemoglobin, and the second dog and cat HbA1c monoclonal antibody can be specifically combined with glycosylated hemoglobin. The glycosylated hemoglobin immunochromatographic test strip provided by the invention achieves quantitative detection of glycosylated hemoglobin of dogs and cats, and has the characteristics of rapidness and simplicity.

By way of summary, the present invention meets some or all of the above needs by providing in a first aspect a method comprising contacting red blood cells in a suspension medium containing mannose under conditions such that the concentration of S-Alc glycated hemoglobin is increased to greater than about 6 percent by weight of the hemoglobin in the red blood cells. Preferably the suspension medium is an aqueous suspension medium. In preferred embodiments, the method is performed at about ambient temperature, 18° C. to about 23° C.

The present invention provides a method of measuring a glycated protein in a sample using a redox reaction, by which the glycated protein can be measured accurately with high sensitivity. In order to remove a glycated amino acid present in the sample other than the glycated protein, the glycated amino acid is degraded in advance by causing a fructosyl amino acid oxidase to act thereon, and thereafter, a fructosyl amino acid oxidase is caused to act on the glycated protein in the presence of a tetrazolium compound and sodium azide to cause a redox reaction. The amount of the glycated protein is determined by measuring the redox reaction. As the glycated protein, glycated hemoglobin is preferable.

This invention provides an amadoriase having high substrate specificity to fructosyl valyl histidine. Such amadoriase comprises substitution of one or more amino acid residues at positions corresponding to amino acids selected from the group consisting of position 98, position 259, position 154, position 125, position 261, position 263, position 106, position 103, position 355, position 96, position 66, position 67, position 70, position 100, position 110, position 113, position 114, and position 156 in the amadoriase derived from the genus Coniochaeta. This invention enables accurate measurement of α-fructosyl valyl histidine derived from the β-chain amino terminus in glycated hemoglobin in the presence of ε-fructosyl lysine.

The invention belongs to the technical field of glycated hemoglobin detection, discloses a glycated hemoglobin and hemoglobin detection kit and a detection method based thereon, and particularly relates to a glycated hemoglobin and hemoglobin detection kit based on boric acid affinity chromatography and a detection method based thereon; the kit comprises reagent R1, reagent R2, reagent R3, reagent R4, and glycated hemoglobin-hemoglobin deposited film; the detection method is specifically a method of detecting glycated hemoglobin and hemoglobin at the same time. Compared with the prior art, the glycated hemoglobin and hemoglobin detection kit and the detection method based thereon have the advantages that by using the kit and detection method, it is possible to shorten the detection time, lower blood taking amount, extend shelf life of the kit, improve detection accuracy, and improve mildness of detection conditions.

The device enables measurement of both of blood glucose level and hemoglobin A1c level, being small-sized and thus portable, allowing the measurement of a small amount of a specimen and, being usable in POC. The device comprises: a test-piece-mounting section (1) for detachably mounting test pieces (200) on which the blood specimen is to be spotted; a light-emitting section (2) for emitting light for irradiating the test pieces; a light-receiving section (3) for receiving reflected lights from the test pieces; and an operating section (4) for calculating the blood glucose level and hemoglobin A1c level of the blood specimen on the basis of photometric values obtained from the light-receiving section. For measuring the blood glucose level of the blood specimen, test piece (A) carries a composition reacting with glucose and showing color change. Test piece (B) carries a composition reacting with glycated hemoglobin and showing color change.

The invention relates to a Chinese medicinal preparation for treating diabetes, which comprises the following Chinese medicinal raw materials in weight proportion: 14 to 16g of red-rooted salvia root, 14 to 16g of astragalus root, 14 to 16g of yam, 14 to 16g of vine root, 9 to 11g of Chinese angelica, 9 to 11g of safflower, and 9 to 11g of ginseng. The preparation method comprises the following steps: cleaning the seven medicinal materials; mixing according to proportion; decocting twice with water in an amount of 5 to 8 times the total weight of the medicinal materials, wherein each decoction takes 1 hour; combining decoctions; and filtering and concentrating to obtain fluid extract with specific gravity of 1.15 at 60 DEG C. The Chinese medicament belongs to compound preparation, can remarkably reduce the level of blood sugar and the content of glycolated hemoglobin, improve the glucose tolerance to a certain extent, promote islet cells to secrete insulin, and does not have acute toxication effect; and compared with the current Chinese medicament for treating diabetes on the market, the Chinese medicament has quick glucose lowering speed, and does not have server adverse reactionof Western medicament products.

The purpose of the present invention is to eliminate the effect of abnormal hemoglobin when a blood sample containing abnormal hemoglobin D, abnormal hemoglobin S, or abnormal hemoglobin C is measuredby cation exchange chromatography, and to obtain an sA1c measurement result reflecting a symptom of the test subject providing the blood sample. The abovementioned problem is solved by a method for measuring the ratio (%) of sA1c, characterized by comprising calculating the area of the peak when a peak originating from abnormal hemoglobin D, abnormal hemoglobin S, or abnormal hemoglobin C is identified and measuring the ratio (%) of sA1c corrected using the calculation result.

There are provided a pre-treatment technique for a glycated hemoglobin-containing sample, which is a simple and convenient treatment, is free from problems in storage stability and environmental aspects, and is capable of exposing an epitope sufficiently in a short time; and an method for an immunological assay of glycated hemoglobin using this technique.A method for pre-treating a glycated hemoglobin-containing sample for an immunological assay of glycated hemoglobin, the method includes treating a glycated hemoglobin-containing sample with a pre-treatment solution containing (A) guanidine or a salt thereof and (B) a nonionic surfactant and / or a nitrite.

There is provided a method for specifically determining a glycated β-chain N-terminal of glycated hemoglobin using enzymes without a separation operation, and a determination reagent kit therefor. A protease that cleaves a glycated amino acid and / or a glycated peptide from a glycated β-chain N-terminal without substantially cleaving a glycated amino acid or a glycated peptide from a glycated α-chain N-terminal of glycated hemoglobin or a fragment thereof is screened. The method of specifically determining a glycated β-chain N-terminal of glycated hemoglobin and the determination reagent kit are provided by using the protease obtained by the screening method. According to the present invention, a glycated β-chain N-terminal of glycated hemoglobin can specifically be determined without a separation operation.

The present invention relates to a chip and a method for detecting HbA1c. The detection chip comprises: a substrate; and a biomolecular layer disposed on the substrate, wherein the biomolecular layer comprises a first anti-hemoglobin antibody. The biomolecular layer can bind both the glycosylated hemoglobin and the hemoglobin in the blood sample. The method for detecting HbA1c of the present invention comprises the use of a bio-detection layer that comprises anti-glycosylated hemoglobin antibody and a second hemoglobin antibody with different epitope to differentiate the glycosylated hemoglobin from the total hemoglobin. Thus, the relative amount of glycosylated hemoglobin to hemoglobin can be detected by using the detection chip and the detection method of the present invention.

The invention provides a micro-fluidic chip for detecting glycosylated hemoglobin and a detection method thereof. The micro-fluidic chip is sequentially provided with a cover plate layer, a fluid layer with a hollow channel structure and a substrate layer from top to bottom, and the hollow channel structure and the substrate layer jointly form a sample detection unit. A sample injection through hole and a waste liquid leading-out through hole are respectively formed in the cover plate layer. The sample detection unit comprises one or more sample injection chambers, a dilution chamber, an affinity chromatography channel, a detection chamber and a waste liquid pool. The sample injection through hole is communicated with the sample injection chamber, and the waste liquid leading-out through hole is communicated with the waste liquid pool. Sample introduction, sample dilution and optical and electrical detection are integrated through the micro-fluidic chip; the operation process is greatly simplified, the consumption of samples and reagents is reduced, expensive equipment is not needed, the use cost is largely reduced, the on-site instant detection of glycosylated hemoglobin can be realized, and huge economic value and social value are created.

The invention relates to a test method of glycosylated hemoglobin, comprising the following steps of: by utilizing the specific adsorption of ion resin to hemoglobin, firstly preparing a hemoglobin sample, fully mixing ion resin with the prepared hemoglobin sample evenly at the temperature of 25-40 DEG C to completely adsorb hemoglobin not glycosylated in blood and keeping glycosylated hemoglobin in blood in liquid, then measuring absorbance values of the liquid and whole blood after adsorption treatment, and obtaining the content of glycosylated hemoglobin according to the ratio of the treated liquid to the treated absorbance value. According to the invention, a microcolumn chromatography is changed to a liquid homogeneous phase adsorption method of separation, so that the operation time is greatly shortened, the operation is simple, the labor intensity of operators is reduced, and the time that a patient waits for a result is shortened; and a disposable plastic cuvette or small bottle is used as a reaction vessel, thus reducing errors and improving degree of precision. The application range is wider, the test method and the test kit can be conveniently used in inspection departments, medical wards and basic medical institutions.

A new method for improvement of functions of diabetes islet beta cells is provided by combination of human umbilical cord mesenchymal stem cells and incretin, and the new method is as follows: on the basis of application of the human umbilical cord mesenchymal stem cells for treatment of type-1 and type-2 diabetes, the incretin (GLP-1 (glucagon-like peptide-1) receptor agonist and DPP-4 (dipeptidyl peptidase-4) inhibitor) is combined. The new method can promote islet beta cell proliferation and regeneration, improve the functions of the islet beta cells, reduce blood sugar and glycated hemoglobin, reduce islet alpha hyperplasia and inhibit glucagon secretion, and thus playing a synergy effect.

The invention discloses an aptamer with high specificity and affinity of glycosylated hemoglobin GHb and a preparation method of the aptamer. The nucleotide sequence of the aptamer of the glycosylated hemoglobin GHb comprises any one of sequences from SEQ ID NO: 1 to SEQ ID NO: 4. The preparation method comprises the following steps of: constructing a random sequence oligonucleotide library, preparing a glycosylated hemoglobin-micro magnetic bead composite, initially screening a target material-nucleic acid composite, preparing a secondary single stranded DNA oligonucleotide library, screening and identifying the aptamer of the glycosylated hemoglobin. The prepared aptamer is free from toxicity, small in molecular weight, and liable to synthesize and mark. The aptamer only specifically identifies the glycosylated hemoglobin GHb, and does not identify other homologous non-glycosylated hemoglobin Hb, so that the aptamer can be taken as a molecular beacon for detecting the content of the glycosylated hemoglobin GHb.

The present invention provides a device for simultaneously detecting hemoglobin and glycohemoglobin in blood, comprising a lateral flow assay strip, a laser-induced epifluorescence detection device, and an LED detection device, and a simple method for simultaneously quantifying hemoglobin and glycohemoglobin in blood by an immunological method using the same.