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Method for measuring glycosylated hemoglobin and kit

A technique for glycosylated hemoglobin and hemoglobin, which is applied in the field of detection methods and kits for glycosylated hemoglobin in isolated blood, can solve the problems of high price, large volume, complicated technical operation, etc., and achieve the effect of simple operation and low price

Inactive Publication Date: 2013-03-27
INTEC PROD INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The above methods all need to detect the two indicators of total hemoglobin and glycated hemoglobin respectively, and then calculate the percentage of glycated hemoglobin in the total hemoglobin to get the final result. The direct consequence of the two tests is that the accuracy of the test results is not high, and now The technical operation is complex and must be operated by professionals
In addition, the instruments used in the prior art are expensive and bulky

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] 1. Magnetic beads coated with anti-hemoglobin antibody. Dilute magnetic beads (produced by Merck, with a particle size of 3um and carboxyl groups on the surface) with 0.1M phosphate buffer to 100mg / ml, and adjust the pH to 7.2. Take 100ml of diluted magnetic beads, add 0.1g carbodiimide and 0.1g hydroxysuccinimide, mix well, and incubate at 37°C for 30 minutes. After the incubation is complete, rinse with 0.1M phosphate buffered saline. After washing, resuspend the magnetic beads in 0.1M phosphate buffer, add 10 mg anti-hemoglobin antibody, mix well and incubate for 30 minutes. After the incubation is complete, rinse with 0.1M phosphate buffered saline. Finally resuspended in 100ml 0.1M phosphate buffered saline, ready for use.

[0025] 2. Whole blood sample lysis. Add 100ul of packed red blood cells to 10ml of pure water and lyse to release the hemoglobin in the red blood cells.

[0026] 3. Magnetic beads adsorb hemoglobin. Add 10ul of the lysed sample to 10ul of...

Embodiment 2

[0034] 1. Magnetic beads coated with anti-hemoglobin antibody. Dilute magnetic beads (produced by Merck, with a particle size of 3um and carboxyl groups on the surface) with 0.1M phosphate buffer to 100mg / ml, and adjust the pH to 7.2. Take 100ml of diluted magnetic beads, add 0.1g carbodiimide and 0.1g hydroxysuccinimide, mix well, and incubate at 37°C for 30 minutes. After the incubation is complete, rinse with 0.1M phosphate buffered saline. After washing, resuspend the magnetic beads in 0.1M phosphate buffer, add 10 mg anti-hemoglobin antibody, mix well and incubate for 30 minutes. After the incubation is complete, rinse with 0.1M phosphate buffered saline. Finally resuspended in 100ml 0.1M phosphate buffered saline, ready for use.

[0035] 2. Whole blood sample lysis. Add 100ul of packed red blood cells to 10ml of pure water and lyse to release the hemoglobin in the red blood cells.

[0036] 3. Magnetic beads adsorb hemoglobin. Add 10ul of the lysed sample to 10ul of...

Embodiment 3

[0042] 1. Whole blood sample lysis. Add 100ul of packed red blood cells to 10ml of pure water and lyse to release the hemoglobin in the red blood cells.

[0043] 2. Magnetic beads adsorb hemoglobin. Add 10 ul of the lysed sample to 10 ul of magnetic beads with polystyrene surface (produced by Dyna Corporation), mix well, incubate at 37°C for 2 minutes, and wash with 0.1M phosphate buffer. Finally resuspended in 20ul 0.1M phosphate buffered saline.

[0044] 3. Add the signal. Add 10ul of rhodamine B and anti-glycosylated hemoglobin antibody conjugate solution to the hemoglobin-adsorbed magnetic bead resuspension. Incubate for 2 minutes. Wash with 0.1M phosphate buffered saline after incubation.

[0045] 4. Reading. After washing, put it into a fluorescence detector for detection. The obtained results are brought into the working curve to obtain the percentage of glycosylated hemoglobin in the sample.

[0046] Using the reagents in this example, commercially available bi...

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PUM

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Abstract

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.

Description

technical field [0001] The invention relates to the field of blood detection, more specifically, to a detection method and kit for glycosylated hemoglobin in isolated blood. Background technique [0002] At present, the commonly used detection methods of glycosylated hemoglobin in clinic include: high performance liquid chromatography, immunoassay, enzymatic method, ion exchange chromatography, chemiluminescence method and so on. High-performance liquid chromatography and ion exchange chromatography require the use of large-scale instruments, and the instruments are expensive; immunoassays require the use of a large number of antibodies, and large-scale instruments are required for detection, such as biochemical analyzers; enzymatic methods require the use of a variety of enzymes Only by reacting can the final result be obtained, and the final result also requires large-scale instrument detection. [0003] The above methods all need to detect the two indicators of total hem...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/72
Inventor 江应玲肖江群王保丹钟乾兴曹大烨汪大明
Owner INTEC PROD INC
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