CN106771261A - The preparation method and its quality-control product of glycosylated hemoglobin quality-control product - Google Patents
The preparation method and its quality-control product of glycosylated hemoglobin quality-control product Download PDFInfo
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- CN106771261A CN106771261A CN201710052326.2A CN201710052326A CN106771261A CN 106771261 A CN106771261 A CN 106771261A CN 201710052326 A CN201710052326 A CN 201710052326A CN 106771261 A CN106771261 A CN 106771261A
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Abstract
The present invention provides a kind of preparation method of glycosylated hemoglobin quality-control product, comprises the following steps:The blood sample of ND is gathered, centrifugal treating is carried out, is removed slurry, obtain red blood cell;To alserver's solution is added in red blood cell, red blood cell sample is obtained;Red blood cell sample is slowly added into layering liquid, the red blood cell being layered is centrifuged, Monodispersed has the red blood cell being adapted to it in the cell separation liquid of each density;The red blood cell of layering is sub-packed in different test tubes, to physiological saline is added in each test tube, red blood cell suspension is obtained, the HC for adjusting the red blood cell suspension is 120g/L, the red blood cell suspension that HbA1c values are 3~5% is filtered out, glycosylated hemoglobin quality-control product is obtained.The present invention also provides a kind of quality-control product.Preparation method of the invention can produce low value glycosylated hemoglobin quality-control product in batches.
Description
Technical field
The present invention relates to technical field of medical examination, more particularly to a kind of glycosylated hemoglobin quality-control product preparation method and
Its quality-control product.
Background technology
The method for clinically determining glycosylated hemoglobin (HbA1c) content at present is different according to physicochemical property, can be divided into two
Major class:One class is based on saccharification and both the non-glycated hemoglobin institute electrically charged difference, such as ion-exchange chromatography, electrophoresis;It is another
Class is, such as immunization, affinity chromatography and enzyme process different from the structure of both the non-glycated hemoglobin based on saccharification.Using different surveys
The reagent determined method or same assay method and select different manufacturers to produce, carries out detecting preceding, it is necessary to carry out matter with quality-control product
Amount control or calibration object are calibrated.At present, self-control Fresh Frozen whole blood is the good Quality Control thing or calibration object of HbA1c detections.
However, the concentration of the HbA1c of normal person is between 5%~6%, the acquisition of various concentrations value has randomness, and uncontrollable, no
Beneficial to industrialization;Wherein HbA1c concentration values be 3%~5% clinical red blood cell sample it is very rare, be difficult obtain.
The content of the invention
It is a primary object of the present invention to provide a kind of preparation method of glycosylated hemoglobin quality-control product, it is desirable to provide a kind of
Batch production HbA1c values are the method for 3~5% glycosylated hemoglobin quality-control product.
To achieve the above object, the preparation method of the glycosylated hemoglobin quality-control product that the present invention is provided, it includes following step
Suddenly:
The blood sample of ND is gathered, is centrifuged, gone slurry to process, obtain red blood cell;
To alserver's solution is added in the red blood cell, red blood cell sample is obtained;
Prepare the different cell separation liquid of density, it is descending according to density, by cell separation liquid sequentially add to one from
In heart pipe, obtain being layered liquid;
The red blood cell sample is slowly added into layering liquid, the red blood cell that centrifugation is layered, the cell of each density
Monodispersed has the red blood cell being adapted to it in separating liquid;
The red blood cell of the layering is sub-packed in different test tubes, to physiological saline is added in each test tube, is obtained
Red blood cell suspension, the HC for adjusting the red blood cell suspension is 120g/L, and it is 3~5% to filter out HbA1c values
Red blood cell suspension, obtain glycosylated hemoglobin quality-control product.
Preferably, after filtering out the red blood cell suspension that HbA1c values are 3~5%, the glycosylated hemoglobin quality-control product
Preparation method also includes:To HbA1c values for 3~5% red blood cell suspension adds buffer solution, haemolysis treatment is carried out, obtained
Solution and the erythrocyte membrane that is scattered in the solution of the HbA1c values for 3~5%;Centrifugation, removes erythrocyte membrane;It is to HbA1c values
3~5% solution adds at least one in freeze drying protectant or stabilizer and carries out the treatment of lyophilized or liquid stored frozen,
Obtain the glycosylated hemoglobin quality-control product.
Preferably, it is added with surfactant in the buffer solution.
Preferably, the density range of the cell separation liquid is 1.085~1.127g/mL.
Preferably, the HC of the blood sample is 120~160g/L, the number of red blood cell in unit volume
It is 4 × 1012~5.5 × 1012The concentration of individual/L, HbA1c is 4~6%.
Preferably, after obtaining cell mixture, before alserver's solution is added in the cell mixture, also including right
The step of cell mixture carries out carrying out washing treatment, the carrying out washing treatment is:To 2~8 DEG C of addition in the cell mixture
Phosphate buffer, be centrifuged 2~5 times, and remove supernatant and tunica albuginea layer.
Preferably, the appearance of the red blood cell in the volumetric ratio of the red blood cell of the alserver's solution and the cell mixture
Product is than equal.
Preferably, described that the red blood cell sample is slowly added into layering liquid, the red blood cell that centrifugation is layered is each
In the step of Monodispersed has the red blood cell being adapted to it in the cell separation liquid of density, centrifuging temperature is 0~20 DEG C, and centrifugal force is
1500~4000g.
Preferably, after the red blood cell of the layering is sub-packed in different test tubes, given birth to being added in each test tube
Before reason salt solution, also including washing the red blood cell with phosphate buffer, and the step of remove supernatant.
The present invention also proposes a kind of quality-control product as obtained by the preparation method of described glycosylated hemoglobin quality-control product.
Compared with prior art, the present invention has the advantages that:Technical solution of the present invention gathers ND
Normal person blood, the final glycosylated hemoglobin quality-control product for obtaining comes from healthy red blood, eliminates matrix effect problem,
The measurement accuracy using the analytical instrument of the glycosylated hemoglobin quality-control product is improved, while have also been enlarged the source of raw material;
Alserver's solution is added in red blood cell, the environment of blood is simulated, so as to maintain the physiologically active of red blood cell.The present invention is using non-
Gradient of continuous density centrifugal process obtains the glycosylated hemoglobin quality-control product of different HbA1c values, and method is easy, is convenient for industrialization
Production.Additionally, obtained glycosylated hemoglobin quality-control product is in a liquid state, can eliminate poor between bottle, improve using the quality-control product point
The certainty of measurement of analyzer device.
Specific embodiment
The technical scheme in the embodiment of the present invention will be clearly and completely described below, it is clear that described implementation
Example is only a part of embodiment of the invention, rather than whole embodiments.Based on the embodiment in the present invention, this area is general
The every other embodiment that logical technical staff is obtained under the premise of creative work is not made, belongs to present invention protection
Scope.
In addition, the technical scheme between each embodiment can be combined with each other, but must be with ordinary skill
Personnel can be implemented as basis, when the combination appearance of technical scheme is conflicting or cannot realize it will be understood that this technical side
The combination of case does not exist, not within the protection domain of application claims yet.
The present invention provides a kind of preparation method of glycosylated hemoglobin quality-control product, and it is comprised the following steps:
The blood sample of ND is gathered, is centrifuged, gone slurry to process, obtain red blood cell;
To alserver's solution is added in the red blood cell, red blood cell sample is obtained;
Prepare the different cell separation liquid of density, it is descending according to density, by cell separation liquid sequentially add to one from
In heart pipe, obtain being layered liquid;
The red blood cell sample is slowly added into the top layer of layering liquid, the red blood cell that centrifugation is layered, each density
Cell separation liquid in Monodispersed have the red blood cell being adapted to it;
The red blood cell of the layering is sub-packed in different test tubes, to physiological saline is added in each test tube, is obtained
Red blood cell suspension, the HC for adjusting the red blood cell suspension is 120g/L, and it is 3~5% to filter out HbA1c values
Red blood cell suspension, obtain glycosylated hemoglobin quality-control product.
The present invention is divided the healthy fresh whole blood red blood cell sample centrifugation of normal person using discontinuous density gradient centrifugation method
From being 2~6 different density levels, so as to filter out the red blood cell of different days.And the percentage of glycosylated hemoglobin (HbA1c)
Specific concentration is relevant with the life-span of red blood cell, and the life-span of red blood cell is more long, and the percent concentration of HbA1c is bigger, otherwise concentration is smaller.
The glycosylated hemoglobin quality-control product of different HbA1c values can be obtained accordingly.
Preferably, alserver's solution is to mix with sodium dihydrogen phosphate containing sodium citrate, citric acid, glucose, adenine
The sterile water solution being made.
The healthy blood sample of normal person of the technical solution of the present invention to collecting carries out centrifugal treating, by blood plasma and cell
Separate, cell mixture is obtained after removal blood plasma;Alserver's solution is added in cell mixture, the environment of blood is simulated,
So as to maintain the physiologically active of red blood cell;The layering liquid of the cell separation liquid with different densities gradient is prepared, by red blood cell sample
Originally it is slowly added into layering liquid, it is preferable that red blood cell sample is located at the top layer of layering liquid;Cause that there are different saccharification after centrifugation
The red blood cell layering of HC, is then dispensed, and adds physiological saline, and regulation HC is 120g/L,
The concentration of hemoglobin i.e. in normal human blood, then with the full-automatic glycosylated hemoglobins of Japan Ai Kelai companies HA-8180 point
Analyzer is detected that screening obtains the red blood cell suspension that HbA1c values are 3~5%.The present invention using discontinuous density gradient from
Heart method obtains the glycosylated hemoglobin quality-control product of different HbA1c values, and method is easy, is convenient for industrialized production.
Additionally, obtained glycosylated hemoglobin quality-control product is in a liquid state, can eliminate poor between bottle, improve using the quality-control product
The certainty of measurement of analytical instrument.
After filtering out the red blood cell suspension that HbA1c values are 3~5%, the preparation side of the glycosylated hemoglobin quality-control product
Method also includes:To HbA1c values for 3~5% red blood cell suspension adds buffer solution, haemolysis treatment is carried out, obtaining HbA1c values is
3~5% solution and the erythrocyte membrane for being scattered in the solution;Centrifugation, removes erythrocyte membrane;It is 3~5% to HbA1c values
Solution adds at least one in freeze drying protectant or stabilizer and carries out the treatment of lyophilized or liquid stored frozen, obtains described
Glycosylated hemoglobin quality-control product.
Preferably, freeze drying protectant or stabilizer are trehalose, sucrose, casein or animal serum albumin.
Technical solution of the present invention carries out haemolysis treatment, adds freeze drying protectant or steady to red blood cell suspension addition buffer solution
Determine agent and carry out lyophilized or liquid stored frozen, preservation 7 days is only capable of under the conditions of 2~8 DEG C compared to red blood cell suspension, effectively
The glycosylated hemoglobin Quality Control thing holding time extended after liquid saccharified hemoglobin Quality Control thing or lyophilized redissolution, user is facilitated to make
With.
Surfactant is added with the buffer solution.
Preferably, the surfactant is Triton X-100 (triton x-100), Tween-20 (polyoxyethylene mountains
Pears sugar alcohol acid anhydride monolaurate).
Surfactant is added with the buffer solution of technical solution of the present invention, SURFACTANT ADSORPTION is in Surface of Erythrocytes
And be inserted into erythrocyte membrane, cause the material on erythrocyte membrane to be reset, and then cause erythrocyte membrane to rupture, effectively improve haemolysis
The efficiency for the treatment of.
The density range of the cell separation liquid is 1.085~1.127g/mL.
Preferably, the density of cell separation liquid can for 1.085,1.0893,1.0905,1.092,1.094,1.098,
1.127g/mL.Layering liquid is made up of the cell separation liquid of at least two different densities, it is preferable that layering liquid includes 2,3,4,5,6
Confluent monolayer cells separating liquid.
The cell separation liquid of technical solution of the present invention has and red blood cell identical density range in blood so that different close
The red blood cell of degree is in floating state in the cell separation liquid of corresponding density, is easy to the layering of different densities red blood cell.
The HC of the blood sample be 120~160g/L, in unit volume the number of red blood cell be 4 ×
1012~5.5 × 1012The concentration of individual/L, HbA1c is 4~6%.
The blood of the normal person of technical solution of the present invention collection ND, the final glycosylated hemoglobin matter for obtaining
Control product come from healthy red blood, eliminate matrix effect problem, improve using the analytical instrument of the glycosylated hemoglobin quality-control product
Measurement accuracy, while have also been enlarged the source of raw material.
After obtaining cell mixture, before alserver's solution is added in the cell mixture, also including to described thin
The step of born of the same parents' mixture carries out carrying out washing treatment, the carrying out washing treatment is:To the phosphoric acid that 2~8 DEG C are added in the cell mixture
Salt buffer, is centrifuged 2~5 times, and removes supernatant and tunica albuginea layer.
Before technical solution of the present invention adds alserver's solution in the cell mixture, washed with phosphate buffer
The cell mixture is washed, the impurity for effectively adhering on removal cell.Wherein the temperature of phosphate buffer is 2~8 DEG C, it is ensured that thin
Born of the same parents are in activated state.
The volumetric ratio phase of the red blood cell in the volumetric ratio of the red blood cell of the alserver's solution and the cell mixture
Deng.
The volumetric ratio of the red blood cell of technical solution of the present invention alserver's solution is equal with the cell mixture, simulates blood
Pendular ring border, so as to maintain the physiologically active of red blood cell.
It is described that the red blood cell sample is slowly added into layering liquid, the red blood cell being layered is centrifuged, each density
In the step of Monodispersed has the red blood cell being adapted to it in cell separation liquid, the temperature of centrifugation is 0~20 DEG C, and centrifugal force is 1500
~4000g.
To being added with the cell separation liquid of red blood cell, be placed in refrigerated centrifuge carries out centrifugal treating to technical solution of the present invention,
The stable state of red blood cell is maintained, while so that the red blood cell layering of different days.Centrifugal force be more than 1500g, effectively improve from
The heart rate of departure, less than 4000g, prevents centrifugal force excessive, causes red blood cell damaged.The temperature of centrifugation is 0~20 DEG C, centrifugal force
It is 1500~4000g, each hierarchy of layer is clearly demarcated, and discrete cell number is less between layers, and separating effect is preferable.
After the red blood cell of the layering is sub-packed in different test tubes, in each test tube add physiological saline it
Before, also including washing the red blood cell with phosphate buffer, and the step of remove supernatant.
Technical solution of the present invention is washed after red blood cell is layered and is dispensed with phosphate buffer, removes red blood cell table
The impurity of face attachment.
The present invention also proposes a kind of quality-control product as obtained by the preparation method of described glycosylated hemoglobin quality-control product.
Embodiment one
The healthy blood sample of normal person is collected, wherein:HC is 120g/L, unit volume red blood cell
Number is 4 × 1012Individual/L, HbA1c:5~6%;
By blood sample prior to 5min is centrifuged with 2000r/min in centrifuge, slurry is removed, the PBS for adding 4 DEG C is washed 2 times,
Rotating speed is also 2000r/min.Carefully inhaled with suction pipe after abandoning supernatant and tunica albuginea layer, obtain red blood cell.
In red blood cell add etc. red cell volume than alserver's solution, fully mix after obtain red blood cell sample,
It is standby in 4 DEG C of Refrigerator stores.
4 Percoll cell separation liquid density gradients are set, be followed successively by 1.120,1.1193,1.1180,1.117g/mL,
It is contained in respectively in 4 centrifuge tubes, 4 part of 400 above-mentioned red blood cell sample of μ L is taken respectively and is slowly superimposed on Percoll along centrifugation tube wall
The superiors of cell separation liquid, set centrifuge temperature for 4 DEG C, centrifugal force are 3500g, and 20min is centrifuged, and obtain being divided into 2 layers
Red blood cell.
The red blood cell suction pipe of the different levels in centrifuge tube is finely drawn, respectively loaded in numbered 2 test tubes,
The phosphate buffer of 4 DEG C of often pipe addition, with 1000r/min, is centrifuged 5min in centrifuge, washs 2 times.Then, supernatant is removed
Liquid, physiological saline is added to each in each test tube, obtains red blood cell suspension, surveys HC, and regulation ultimate density is
120g/L。
HbA1c values are detected with the full-automatic glycolated hemoglobin analysis of Japanese Ai Kelai companies HA-8180, the superiors are chosen
Red blood cell as sample, by the density of the liquid-tight degree of Percoll cell separations be 1.120,1.1193,1.1180,1.117g/mL
Treatment obtains low HbA1c values sample number for 1,2,3,4 groups respectively, wherein undressed original blood sample is No. 0.By table
When 1 density that can be seen that the liquid-tight degree of Percoll cell separations is 1.117g/mL, can be relatively low with isolated HbA1c values
Red blood cell.
The different experiments group upper strata red blood cell HA-8180 testing results of table 1
Group | HbA1c values | With original value bias |
0 | 5.3% | —— |
1 | 5.3% | 0% |
2 | 5.3% | 0% |
3 | 5.2% | - 0.1% |
4 | 4.8% | - 0.5% |
The buffer solution containing surfactant is added in red blood cell after separating the layers, haemolysis treatment is carried out, is removed with centrifugation
Impurity, after adding freeze drying protectant/stabilizer, freezes or liquid stored frozen.
Embodiment two
The healthy blood sample of normal person is collected, wherein:HC is 120g/L, unit volume red blood cell
Number is 4 × 1012Individual/L, HbA1c:5~6%;
By blood sample prior to 5min is centrifuged with 2000r/min in centrifuge, slurry is removed, the PBS for adding 2 DEG C is washed 5 times,
Rotating speed is also 2000r/min.Carefully inhaled with suction pipe after abandoning supernatant and tunica albuginea layer, obtain red blood cell.
In red blood cell add etc. red cell volume than alserver's solution, fully mix after obtain red blood cell sample,
It is standby in 4 DEG C of Refrigerator stores.
Percoll cell separations stoste is a kind of hypo-osmoticity medium, need to add the NaCl of 1.5mol/L molten when being prepared
Liquid, can just make the osmotic pressure of the Percoll cell separation liquid after preparation reach the required size (280- of red blood cell
320mOsm/kg·H2O)。
According to equation below:
V0The volume of-undiluted Percoll cell separations liquid stoste, unit is mL
The volume of the Percoll cell separation liquid after V-dilution, unit is mL
ρ0The density of-undiluted Percoll cell separations liquid, is 1.130g/mL
The density of the Percoll cell separation liquid after ρ-dilution
ρ10The density of the NaCl solution of -1.5mol/L, is 1.058g/mL
Calculate 3 kinds of liquid when preparing different densities separating liquid (Percoll cell separation liquid stoste, 0.9% physiological saline,
The NaCl solution of 1.5mol/L) proportioning.
6 kinds are prepared with Percoll cell separation liquid stostes and 0.9% physiological saline, the NaCl solution of 1.5mol/L not
With the separating liquid of density, density from low to high be respectively 1.085,1.0893,1.0905,1.092,1.094,1.098g/mL (according to
Secondary to be designated as 1,2,3,4,5,6,0) untreated whole blood is designated as.Above-mentioned separating liquid is descending according to density, it is added sequentially to
In one centrifuge tube, finally red blood cell sample is added in centrifuge tube, now red blood cell sample is located at the top layer of layering liquid.
Centrifugal condition be set to 3500g, 10 DEG C, be centrifuged 20min, red blood cell be located at corresponding density cell separation liquid in.
The red blood cell suction pipe of the different levels in centrifuge tube after centrifugation is finely drawn, respectively loaded in numbered 6 test tubes, is gone
Supernatant, to the physiological saline of each addition 1mL in each pipe, is obtained red blood cell suspension, adjusts the blood red egg of red blood cell suspension
White concentration is 120g/L.
Detected with the Japanese full-automatic glycolated hemoglobin analysis of Ai Kelai companies HA-8180, measure this six layers it is red
The HbA1c values of cell, as shown in table 2, are followed successively by from top to bottom:3.5%th, 3.8%, 4.2%, 4.5%, 4.8%, 5.3%.Say
Bright use the method can obtain 6 products of HbA1c levels, wherein HbA1c values be 3.5%, 3.8%, 4.2%, 4.5%,
4.8% red blood cell suspension, is comparatively ideal low-saccharification hemoglobin quality-control product.
The different layers red blood cell HA-8180 testing results of table 2
Layer is not | HbA1c values | With original value bias |
0 | 5.2% | —— |
1 | 3.5% | - 1.7% |
2 | 3.8% | - 1.4% |
3 | 4.2% | - 1.0% |
4 | 4.5% | - 0.7% |
5 | 4.8% | - 0.4% |
6 | 5.3% | 0.1% |
By above-mentioned HbA1c values for 3.5%, 3.8%, 4.2%, 4.5%, 4.8% red blood cell suspension is positioned over refrigerator
In it is standby, can be preserved under the conditions of 2~8 DEG C 7 days.
Using the NaCl solution of be added with Triton X-100 0.05%, haemolysis treatment is carried out to red blood cell suspension,
The removal of impurity is gone in centrifugation, to animal serum albumin is added in the solution that centrifugation is obtained, carries out liquid frost treatment, the saccharification for obtaining
Hemoglobin quality-control product can be preserved 3 months.
Embodiment three
The healthy blood sample of normal person is collected, wherein:HC is 160g/L, unit volume red blood cell
Number is 5.5 × 1012Individual/L, HbA1c:6%;
By blood sample prior to 5min is centrifuged with 2000r/min in centrifuge, slurry is removed, the PBS for adding 8 DEG C is washed 5 times,
Rotating speed is also 2000r/min.Carefully inhaled with suction pipe after abandoning supernatant and tunica albuginea layer, obtain red blood cell.
In red blood cell add etc. red cell volume than alserver's solution, fully mix after obtain red blood cell sample,
It is standby in 4 DEG C of Refrigerator stores.
6 kinds are prepared with Percoll cell separation liquid stostes and 0.9% physiological saline, the NaCl solution of 1.5mol/L not
With the separating liquid of density, density from low to high be respectively 1.085,1.0893,1.0905,1.092,1.094,1.098g/mL (according to
Secondary to be designated as 1,2,3,4,5,6,0) untreated whole blood is designated as.Above-mentioned separating liquid is descending according to density, it is added sequentially to
In one centrifuge tube, finally red blood cell sample is added in centrifuge tube, now red blood cell sample is located at the top layer of layering liquid.
Centrifugal condition be set to 4000g, 20 DEG C, be centrifuged 20min, red blood cell be located at corresponding density cell separation liquid in.
The red blood cell suction pipe of the different levels in centrifuge tube after centrifugation is finely drawn, respectively loaded in numbered 6 test tubes, is gone
Supernatant, to the physiological saline of each addition 1mL in each pipe, is obtained red blood cell suspension, adjusts the blood red egg of red blood cell suspension
White concentration is 120g/L.Detection HbA1c is carried out with the Japanese full-automatic glycolated hemoglobin analysis of Ai Kelai companies HA-8180
Value, filters out the red blood cell suspension that HbA1c values are 3~5%, is saved backup under the conditions of 2~8 DEG C.
The preferred embodiments of the present invention are these are only, the scope of the claims of the invention is not thereby limited, it is every to utilize this hair
Equivalent structure or equivalent flow conversion that bright specification is made, or other related technical fields are directly or indirectly used in,
Similarly it is included within the scope of the present invention.
Claims (10)
1. a kind of preparation method of glycosylated hemoglobin quality-control product, it is comprised the following steps:
The blood sample of ND is gathered, is centrifuged, gone slurry to process, obtain red blood cell;
To alserver's solution is added in the red blood cell, red blood cell sample is obtained;
The different cell separation liquid of density is prepared, it is descending according to density, cell separation liquid is sequentially added to a centrifuge tube
It is interior, obtain being layered liquid;
The red blood cell sample is slowly added into layering liquid, the red blood cell that centrifugation is layered, the cell separation of each density
Monodispersed has the red blood cell being adapted to it in liquid;
The red blood cell of the layering is sub-packed in different test tubes, to physiological saline is added in each test tube, obtains red thin
Born of the same parents' suspension, the HC for adjusting the red blood cell suspension is 120g/L, filter out HbA1c values be 3~5% it is red
Cell suspending liquid, obtains glycosylated hemoglobin quality-control product.
2. the preparation method of glycosylated hemoglobin quality-control product as claimed in claim 1, it is characterised in that filter out HbA1c values
After for 3~5% red blood cell suspension, the preparation method of the glycosylated hemoglobin quality-control product also includes:It is 3 to HbA1c values
~5% red blood cell suspension addition buffer solution, carries out haemolysis treatment, obtains solution and be scattered in that HbA1c values are 3~5%
The erythrocyte membrane of the solution;Centrifugation, removes erythrocyte membrane;To HbA1c values for 3~5% solution add freeze drying protectant or
At least one in stabilizer simultaneously carries out the treatment of lyophilized or liquid stored frozen, obtains the glycosylated hemoglobin quality-control product.
3. the preparation method of glycosylated hemoglobin quality-control product as claimed in claim 2, it is characterised in that add in the buffer solution
Added with surfactant.
4. the preparation method of glycosylated hemoglobin quality-control product as claimed in claim 1, it is characterised in that the cell separation liquid
Density range be 1.085~1.127g/mL.
5. the preparation method of glycosylated hemoglobin quality-control product as claimed in claim 1, it is characterised in that the blood sample
HC is 120~160g/L, and the number of red blood cell is 4 × 10 in unit volume12~5.5 × 1012Individual/L, HbA1c
Concentration be 4~6%.
6. the preparation method of glycosylated hemoglobin quality-control product as claimed in claim 1, it is characterised in that obtain cell mixture
Afterwards, before to alserver's solution is added in the cell mixture, also including carrying out carrying out washing treatment to the cell mixture
Step, the carrying out washing treatment is:To the phosphate buffer that 2~8 DEG C are added in the cell mixture, it is centrifuged 2~5 times, and
Removal supernatant and tunica albuginea layer.
7. the preparation method of glycosylated hemoglobin quality-control product as claimed in claim 1, it is characterised in that the red cell preservation
The volumetric ratio of the red blood cell of liquid is equal with the volumetric ratio of the red blood cell in the cell mixture.
8. the preparation method of glycosylated hemoglobin quality-control product as claimed in claim 1, it is characterised in that it is described will be described red thin
Born of the same parents' sample is slowly added into layering liquid, the red blood cell that centrifugation is layered, in the cell separation liquid of each density Monodispersed have with
In the step of red blood cell of its adaptation, centrifuging temperature is 0~20 DEG C, and centrifugal force is 1500~4000g.
9. the preparation method of glycosylated hemoglobin quality-control product as claimed in claim 1, it is characterised in that by the layering
After red blood cell is sub-packed in different test tubes, to before adding physiological saline in each test tube, also including using phosphate-buffered
Liquid washs the red blood cell, and the step of remove supernatant.
10. the matter obtained by a kind of preparation method of glycosylated hemoglobin quality-control product as any one of claim 1-9
Control product.
Priority Applications (1)
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CN110806450B (en) * | 2019-11-14 | 2022-03-25 | 广州科方生物技术股份有限公司 | Liquid calibrator for glycosylated hemoglobin |
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