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Preparation method of glycosylated hemoglobin control product and quality control product thereof

A hemoglobin and saccharification technology, applied in the field of medical testing, can solve problems such as unfavorable industrialization, difficult to obtain, uncontrollable, etc., and achieve the effects of improving measurement accuracy, simple method, and eliminating bottle-to-bottle difference.

Active Publication Date: 2019-05-24
深圳市雷诺华科技实业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the HbA1c concentration of normal people is between 5% and 6%, and the acquisition of different concentration values ​​is random and uncontrollable, which is not conducive to industrialization; clinical red blood cell samples with an HbA1c concentration of 3% to 5% are very rare, not easy to get

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  • Preparation method of glycosylated hemoglobin control product and quality control product thereof

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preparation example Construction

[0022] The invention provides a method for preparing a glycosylated hemoglobin control product, which comprises the following steps:

[0023] Collect blood samples from non-diabetic patients, centrifuge and deslurry to obtain red blood cells;

[0024] adding a red blood cell preservation solution to the red blood cells to prepare a red blood cell sample;

[0025] Prepare cell separation liquids with different densities, and add the cell separation liquids to a centrifuge tube sequentially according to the density from large to small to obtain a layered liquid;

[0026] The red blood cell sample is slowly added to the surface layer of the layered liquid, and centrifuged to obtain layered red blood cells, and the red blood cells adapted to each density are dispersed in the cell separation liquid;

[0027] The layered erythrocytes were subpackaged in different test tubes, and physiological saline was added to each test tube to obtain a erythrocyte suspension, and the hemoglobin ...

Embodiment 1

[0053] Collect healthy blood samples from normal people, in which: the hemoglobin concentration is 120g / L, and the number of red blood cells per unit volume is 4×10 12 A / L, HbA1c: 5-6%;

[0054] The blood sample was first centrifuged in a centrifuge at 2000r / min for 5min to remove the pulp, and then washed twice with PBS at 4°C at a rotational speed of 2000r / min. After the supernatant and buffy coat were carefully discarded with a pipette, red blood cells were obtained.

[0055] Add erythrocyte preservation solution with equal erythrocyte volume ratio to erythrocytes, mix thoroughly to obtain erythrocyte samples, and store them in a refrigerator at 4°C for later use.

[0056] Set up 4 density gradients of Percoll cell separation medium, which are 1.120, 1.1193, 1.1180, and 1.117g / mL in sequence, and put them in 4 centrifuge tubes respectively. Take 4 parts of 400 μL of the above red blood cell samples and slowly superimpose them on the Percoll cell separation medium along the...

Embodiment 2

[0063] Collect healthy blood samples from normal people, in which: the hemoglobin concentration is 120g / L, and the number of red blood cells per unit volume is 4×10 12 A / L, HbA1c: 5-6%;

[0064] The blood sample was first centrifuged in a centrifuge at 2000r / min for 5min, deslurried, and then washed 5 times with PBS at 2°C, and the rotation speed was also 2000r / min. After the supernatant and buffy coat were carefully discarded with a pipette, red blood cells were obtained.

[0065] Add erythrocyte preservation solution with equal erythrocyte volume ratio to erythrocytes, mix thoroughly to obtain erythrocyte samples, and store them in a refrigerator at 4°C for later use.

[0066] Percoll cell separation stock solution is a hypotonic medium. When preparing, it is necessary to add 1.5mol / L NaCl solution to make the osmotic pressure of the prepared Percoll cell separation solution reach the required size of red blood cells (280-320mOsm / kg ·H 2 O).

[0067] According to the fol...

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Abstract

The invention provides a preparation method of a quality control material of glycosylated hemoglobin, comprising the following steps: collecting a blood sample of a nondiabetic patient, performing centrifugation and plasmapheresis to obtain red blood cells; adding red blood cell preserving fluid to the red blood cells to obtain a red blood cell sample; adding the red blood cell sample slowly into separating fluid and centrifuging to obtain separated red blood cells, wherein red blood cells adaptive to the cell separation fluid are dispersed in the cell separation fluid of each density; separately charging the separated red blood cells into different test tubes, adding physiological saline to each test tube to obtain a red blood cell suspension, regulating the hemoglobin concentration of the red blood cell suspensions to 120g / L, and screening out red blood cell suspensions with the HbA1c value of 3-5% to obtain the quality control material of glycosylated hemoglobin. The invention also provides the quality control material. The preparation method provided by the invention can perform mass production of low-value quality control materials of glycosylated hemoglobin.

Description

technical field [0001] The invention relates to the technical field of medical testing, in particular to a method for preparing a glycosylated hemoglobin control product and a quality control product thereof. Background technique [0002] At present, the clinical methods for determining the content of glycated hemoglobin (HbA1c) can be divided into two categories according to different physical and chemical properties: one is based on the difference in charge between glycated and non-glycated hemoglobin, such as ion exchange chromatography and electrophoresis; It is based on the different structures of glycated and non-glycated hemoglobin, such as immunoassay, affinity chromatography and enzymatic method. If different assay methods or reagents produced by different manufacturers are selected for the same assay method, quality control products or calibrator are required for quality control or calibration before testing. At present, self-made fresh frozen whole blood is a goo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/72
CPCG01N33/723G01N2333/805G01N2496/05
Inventor 王荣光
Owner 深圳市雷诺华科技实业有限公司
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