CN103454410B - Biologically-safe quality control material for blood tester and preparation method thereof - Google Patents
Biologically-safe quality control material for blood tester and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a biologically-safe quality control material for a blood tester and a preparation method thereof. The preparation method comprises that animal red blood cells having the sizes similar to sizes of human red blood cells are used for simulation of human red blood cells; animal red blood cells having the sizes similar to sizes of large cells in human white blood cells and animal red blood cells have the sizes similar to sizes of small cells in human white blood cells are used for simulation of human white blood cells; animal red blood cells have the sizes similar to sizes of human blood platelets are used for simulation of human blood platelets; the animal red blood cells for simulation of human white blood cells and human blood platelets are cured by formaldehyde having the content of 2-8%; the cured animal red blood cells are cleaned and then are added with bovine serum albumin so that the cure agent is removed; and according to requirements, through blending, the single-component or whole blood quality control material is obtained. The biologically-safe quality control material has good stability, a low cost and high safety, and is suitable for a blood cell analyzer, a urinary sediment analyzer and a hemoglobin analyzer.
Description
Technical field
The present invention relates to blood testing technical field, be specifically related to Biologically-safe quality control material for blood tester and preparation method thereof.
Background technology
Quality Control thing is used for the performance of Inspection and analysis instrument or method.When material is used for the conventionally test of Inspection and analysis instrument or method current performance, its analytical characteristics must similar to clinical samples.
Along with the develop rapidly of medical diagnostic techniqu, the use of various blood clinical testing tool is more and more advanced and general, not only provides more experimental index for clinical, also substantially increases clinical trial work efficiency and accuracy.But result accurately and reliably will be obtained, the result that different instrument and different experiments room are detected has comparability, needs a kind of Quality Control thing that is stable, reliable, multiparameter normal to confirm the operating function running of instrument, every location parameter whether bias.The large person of Quality Control thing that current domestic various big hospital uses adopts external Quality Control thing, and external Quality Control owner will adopt human red cell, birds karyocyte, latex, pollen, hoof section animal erythrocyte, makes the red blood cell of human red cell simulation Quality Control thing.Costly, and owing to being adopt human red cell to make Quality Control thing, thus likely there is the problem of blood stains dye, also may cause other transmissions of disease by blood-borne such as hepatitis, acquired immune deficiency syndrome (AIDS), venereal disease in external its price of Quality Control thing.At present; domestic also have the similar quality-control product of human blood; the patent of invention being CN101285841 as publication number discloses a kind of preparation method of three classifications whole blood quality control substance simulant and obtained blood type detection kit thereof; the described method Fish Blood simulation human leukocyte of this invention and blood platelet; domestic animal red blood cell and slurry station discarded human red cell simulation human red cell; the histogram distribution of simulation human blood sample reality; cell granulations is suspended in cell suspension and does the use of individual event Quality Control thing, or be combined into whole blood quality control materials use.This Quality Control thing of the present invention has good Quality Control effect, but its pot-life is not ideal enough.
Summary of the invention
The technical problem to be solved in the present invention is to provide one and adopts animal blood to make completely, reach bio-safety and Quality Control is effective while storage life limit for length blood tester Quality Control thing and preparation method thereof.
The preparation method of blood tester Quality Control thing of the present invention, comprises the preparation of (1) red blood cell analogies and the preparation of (2) leucocyte and platelet analogue, wherein:
(1) preparation method of described red blood cell analogies is as follows:
A) gather red blood cell be similar to the blood of the animal of human red cell size and adopt anti-coagulants process;
B) by the above-mentioned centrifugal blood collected, remove blood plasma and leucocyte, collect bottom red blood cell;
C) red blood cell of collection is placed in cell maintenance liquid to preserve, namely obtains red blood cell analogies;
(2) preparation method of leucocyte and platelet analogue is as follows:
D) gather the blood that the blood that red blood cell is similar to the blood of the Magnocellular animal of human blood leukocyte, red blood cell is similar to the animal of human blood leukocyte's cellule and red blood cell are similar to the animal of human blood Platelet Size respectively, while collection, carry out anti-coagulants process;
E) be cured respectively by the above-mentioned blood collected, concrete curing operation is: by the centrifugal blood collected, and collect the red blood cell of bottom, the formaldehyde adding 2 ~ 8% (v/v) is cured 12 ~ 72h;
F) blood after above-mentioned three kinds of solidifications is carried out the step removing hardening agent respectively, the operation of concrete removal hardening agent be by solidification after centrifugal blood, remove supernatant, retain cell to clean with physiological saline, cell after cleaning adds bovine serum albumin, leave standstill 20 ~ 24h, centrifugal, isolate bottom cell;
The red blood cell of the above-mentioned isolated human blood leukocyte's of being similar to maxicell animal is placed in cell maintenance liquid preserve, namely obtains leucocyte maxicell analogies; The red blood cell of the above-mentioned isolated human blood leukocyte's of being similar to cellule animal is placed in cell maintenance liquid preserve, namely obtains leucocyte cellule analogies; The isolated red blood cell being similar to the animal of human blood platelets is placed in cell maintenance liquid preserve, namely obtains platelet analogue;
G) regulate the concentration of red blood cell analogies and platelet analogue to can be used as individual event Quality Control thing respectively to use, or leucocyte maxicell analogies and leucocyte cellule analogies are mixed to get leucocyte analogies use as individual event Quality Control thing; Or the combination of described red blood cell analogies, leucocyte maxicell analogies, leucocyte cellule analogies and platelet analogue is used as whole blood quality control materials;
In above-mentioned preparation method, the osmotic pressure of the cell maintenance liquid mentioned is 300 ± 20mOsm/kg, pH value is 7.3 ± 0.1, and it consists of: microbiotic 0.4 ~ 2g/L, polysaccharide 5 ~ 20g/L, bovine serum albumin 20 ~ 50g/L, sodium chloride 0.1 ~ 10g/L, sodium citrate 0.1 ~ 20g/L and citric acid 0.001 ~ 5g/L.
In technique scheme:
Steps A) in, the animal that described red blood cell is similar to human red cell size can be cat, dog, pig, rabbit, monkey, ox, chicken or duck.
Step D) in, it can be goose, wild goose, tortoise, ostrich, snake, shark or crocodile that described red blood cell is similar to the Magnocellular animal of human blood leukocyte.
Step D) in, the animal that described red blood cell is similar to human blood leukocyte's cellule can be cat, dog, pig, rabbit, monkey, ox, chicken or duck.
Step D) in, the animal that described red blood cell is similar to human blood Platelet Size can be cat, horse, ox, sheep or camel.
Step e) in, the addition of formaldehyde is preferably 40:1 ~ 1:1 with the erythrocytic volume ratio collected.
Microbiotic in described cell maintenance liquid is selected from penicillins, aminoglycoside and amide-type antibiotic any one or two or more combinations.Can be specifically adopt one or more the combination in penicillin, tardocillin, gentamicin, streptomysin and chloromycetin.When antibiotic be chosen as above-mentioned two or more combination time, the proportioning between them can be any proportioning.
Polysaccharide in described cell maintenance liquid is selected from the combination of any one or more in sucrose, glucose, fructose, galactose, lactose, maltose, starch and dextrin.When polysaccharide be chosen as above-mentioned two or more combination time, the proportioning between them can be any proportioning.
Described anti-coagulants is the conventional anticoagulants adopted in conventional blood collection.
The present invention also comprises the Quality Control thing prepared by above-mentioned preparation method.
Compared with prior art, the invention has the advantages that:
1, only need to be cured the animal erythrocyte of simulation human leukocyte and human blood platelets, simplify technique;
2, adopt the animal erythrocyte of the formaldehyde of high concentration to simulation human leukocyte and human blood platelets to be cured, high-concentration formaldehyde energy rapid curing cell, solidification is more thorough, shows from stability observing, and solidification concentration is high, can ensure that solidification cell stability is longer; After hardening by cleaning and adding the residual hardening agent of bovine serum albumin removing, overcome and cause cell volume in finished product Quality Control thing to change because solidifying to remain, ensure that the stability of Quality Control thing;
3, adopt the cell maintenance liquid of special proportioning, ensure erythrocytic activity and stability;
4, all adopt animal blood to make, cost is lower, and security is higher;
5, Quality Control thing adopts animal blood to make, and can stop the harm that people's blood borne diseases uses at product, carry out work under allowing user can be in better environment; And raw material sources are extensive, avoid the contention of people's blood resource, it is convenient to gather, and controllability is large, greatly increases product practicality, increases the diversity of domestic Quality Control produce product, promotes the benign competition of market like product;
6, Quality Control thing storage life limit for length of the present invention, applied widely, is applicable to cellanalyzer, urine sediments analyzer and hemoglobin analyser.
Embodiment
With embodiment, end invention is further described below, but the present invention is not limited to these embodiments.
Embodiment 1: the preparation of the Quality Control thing of hemoglobin analyser
A) gather blood of goats and carry out anti-freezing process, anti-coagulants is EDTA-K2, and consumption is that 1.5 ~ 2.2MG/ml blood carries out anti-freezing process;
B) by centrifugal for the above-mentioned blood of goats collected, remove blood plasma and leucocyte, collect bottom red blood cell, obtain concentration higher than 95% red blood cell;
C) red blood cell of collection is placed in cell maintenance liquid, according to the content of haemoglobin in the Quality Control thing that will prepare, regulates the concentration of red blood cell in cell maintenance liquid, namely obtain the haemoglobin Quality Control thing of respective concentration; Consisting of of cell maintenance liquid: penicillin 0.15g/L, gentamicin 0.15g/L, chloromycetin 0.1g/L, glucose 2g/L, bovine serum albumin 20g/L, sodium chloride 6.3g/L, sodium citrate 7.8g/L, citric acid 0.01g/L; The osmotic pressure of described cell maintenance liquid is 305mOsm/kg, and pH value is 7.32.
The preparation of the whole blood quality control materials of embodiment 2: three classification cellanalyzer
A) gather the blood of dog, ox and ostrich respectively, in the process gathered, use anti-coagulants process respectively; EDTA-K selected by anti-coagulants
2, consumption is 2.0MG/ml blood;
A part of B) getting the dog blood of collection is carried out centrifugal, removes upper plasma and middle level leucocyte, collects bottom red blood cell, obtains the red blood cell that concentration is 98.5%;
C) red blood cell collected is mixed with Precerving liquid, with cellanalyzer test, be adjusted to RBC with Precerving liquid and be counted as 8.0X10
12/ L ± 1.0X10
12/ L, volume ratio is about 1:1, preserves, namely obtain red blood cell analogies under 5 DEG C of conditions;
D) by centrifugal for remaining dog blood, collect the red blood cell of bottom, the formaldehyde adding 3% (v/v) solidifies 56h in the constant temperature oven of 30 DEG C; The addition of described formaldehyde is 20:1 with the erythrocytic volume ratio collected;
The ox blood getting above-mentioned collection is centrifugal, and collect the red blood cell of bottom, the formaldehyde adding 3% (v/v) solidifies 56h in the constant temperature oven of 30 DEG C; The addition of described formaldehyde is 20:1 with the erythrocytic volume ratio collected;
The ostrich blood getting above-mentioned collection is centrifugal, and collect the red blood cell of bottom, the formaldehyde adding 3% (v/v) solidifies 56h in the constant temperature oven of 30 DEG C; The addition of described formaldehyde is 20:1 with the erythrocytic volume ratio collected;
E) the dog blood after above-mentioned solidification is carried out the operation of removing hardening agent, specifically by centrifugal for the dog blood after solidification, remove supernatant, retain cell then centrifugal with physiological saline cleaning, repeat 3 cleaning operations, it is 38% bovine serum albumin (wherein the volume ratio of cell and bovine serum albumin is 1:1) that the centrifugal cell obtained adds concentration, centrifugal after leaving standstill 24h, isolated bottom cell is the dog red blood cell (concentration is 98%) being similar to human blood leukocyte's cellule, it is mixed with the volume ratio of cell maintenance liquid by 1:2, namely obtains SL analogies;
Ox blood after above-mentioned solidification is carried out the operation of removing hardening agent, concrete operations are with dog blood, isolated bottom cell is the ORBC (concentration is 99%) being similar to human blood platelets, it is mixed with the volume ratio of cell maintenance liquid by 1:3, namely obtains platelet cell analogies;
Ostrich blood after above-mentioned solidification is carried out the operation of removing hardening agent, concrete operations are with dog blood, isolated bottom cell is and is similar to the Magnocellular ostrich red blood cell of human blood leukocyte (concentration is 98.5%), it is mixed with the volume ratio of cell maintenance liquid by 1:5, namely obtains large leucocyte analogies;
F) by step C) in obtained red blood cell analogies, step e) obtained leucocyte cellule analogies, platelet analogue and the leucocyte maxicell analogies volume ratio of press 3:1:1:1 mixes, packing, namely obtains the whole blood quality control materials of three classification cellanalyzers;
Consisting of of the cell maintenance liquid mentioned in said method: penicillin 0.2g/L, gentamicin 0.25g/L, chloromycetin 0.1g/L, glucose 2.5g/L, fructose 1.5g/L, galactose 2g/Lg/L, bovine serum albumin 25g/L, sodium chloride 6g/L, sodium citrate 8.0g/L, citric acid 0.01g/L; Its osmotic pressure is 300mOsm/kg, and pH value is 7.35.
By Quality Control thing obtained for above-described embodiment 2, observe homogeneity in homogeneity, bottle in the bottle of Quality Control thing and accuracy with cellanalyzer, result is as following:
1, test result
Homogeneity in 1.1 bottles
Get arbitrarily Quality Control thing 1 bottle, get 3 samples, be numbered sample 1, sample 2 and sample 3, test with cellanalyzer URIT-3000 (Guilin High-Tech Zone Baolitai Medical Electronic Co., Ltd.) respectively, repeated test 11 times, remove the 1st test result, get residue 10 test results, as described in Table 1.Intermediate value Quality Control thing parameters presses the homogeneity coefficient of variation in formula (1), formula (2), formula (3) calculating bottle, and result is as shown in table 2, meets industry standard.
In formula:
--mean value;
SD--standard deviation;
The CV--coefficient of variation;
N--pendulous frequency;
X
i--designated parameter i-th measured value.
Table 1:
Table 2:
Homogeneity between 1.2 bottles
Get 10 bottles of Quality Control things of the same lot number prepared by embodiment 2 method, every bottle of Quality Control thing tests 2 times, removes the 1st test result, calculates the mean value of parameters 10 results by formula (1), formula (2)
with standard deviation (SD
1);
Separately with 1 bottle of follow-on test in above-mentioned 10 bottles of Quality Control things 11 times, remove the 1st test result, by the mean value of formula (1), formula (2) calculating residue 10 test result parameters
with standard deviation (SD
2);
Intermediate value Quality Control thing parameters presses formula (4), formula (5) calculates homogeneity between bottle, and between gained bottle, homogeneity coefficient of variation result is as shown in table 3, meets industry standard.
Work as SD
1< SD
2time, make CV
between bottle=0
In formula:
---mean value;
SD---standard deviation.
Table 3:
2, accuracy
Producer's assignment
| Parameter | WBC×10 9/L | RBC×10 12/L | HGB g/L | MCV fL | PLT×10 9/L |
| With reference to assignment | 9.6±0.6 | 4.43±0.2 | 86±5 | 61.3±5 | 237±30 |
Get by the obtained Quality Control thing of embodiment 2 method, carry out trimestral observation test by a definite date, as described in Table 4, Quality Control thing long-term test accuracy result is as shown in table 5 for determination data.
Table 4:
Table 5:
As can be seen from above-mentioned every observation test, the Quality Control thing made by the method for the invention meets the requirement of homogeneity and the accuracy of Quality Control thing between homogeneity in bottle, bottle.
Claims (6)
1. the preparation method of Biologically-safe quality control material for blood tester, comprises the preparation of (1) red blood cell analogies and the preparation of (2) leucocyte and platelet analogue, it is characterized in that:
(1) preparation method of described red blood cell analogies is as follows:
A) gather red blood cell be similar to the blood of the animal of human red cell size and adopt anti-coagulants process;
B) by the above-mentioned centrifugal blood collected, remove blood plasma and leucocyte, collect bottom red blood cell;
C) red blood cell of collection is placed in cell maintenance liquid to preserve, namely obtains red blood cell analogies;
(2) preparation method of leucocyte and platelet analogue is as follows:
D) gather the blood that the blood that red blood cell is similar to the blood of the Magnocellular animal of human blood leukocyte, red blood cell is similar to the animal of human blood leukocyte's cellule and red blood cell are similar to the animal of human blood Platelet Size respectively, while collection, carry out anti-coagulants process;
E) be cured respectively by the above-mentioned blood collected, concrete curing operation is: by the centrifugal blood collected, and collect the red blood cell of bottom, the formaldehyde adding 2 ~ 8% (v/v) is cured 12 ~ 72h;
F) blood after above-mentioned three kinds of solidifications is carried out the step removing hardening agent respectively, the operation of concrete removal hardening agent be by solidification after centrifugal blood, remove supernatant, retain cell to clean with physiological saline, cell after cleaning adds bovine serum albumin, leave standstill 20 ~ 24h, centrifugal, isolate bottom cell;
The red blood cell of the above-mentioned isolated human blood leukocyte's of being similar to maxicell animal is placed in cell maintenance liquid preserve, namely obtains leucocyte maxicell analogies; The red blood cell of the above-mentioned isolated human blood leukocyte's of being similar to cellule animal is placed in cell maintenance liquid preserve, namely obtains leucocyte cellule analogies; The isolated red blood cell being similar to the animal of human blood platelets is placed in cell maintenance liquid preserve, namely obtains platelet analogue;
G) regulate the concentration of red blood cell analogies and platelet analogue to can be used as individual event Quality Control thing respectively to use, or leucocyte maxicell analogies and leucocyte cellule analogies are mixed to get leucocyte analogies use as individual event Quality Control thing; Or the combination of described red blood cell analogies, leucocyte maxicell analogies, leucocyte cellule analogies and platelet analogue is used as whole blood quality control materials;
In above-mentioned preparation method, the osmotic pressure of the cell maintenance liquid mentioned is 300 ± 20mOsm/kg, pH value is 7.3 ± 0.1, and it consists of: microbiotic 0.4 ~ 2g/L, polysaccharide 0.1 ~ 20g/L, bovine serum albumin 20 ~ 50g/L, sodium chloride 0.1 ~ 10g/L, sodium citrate 0.1 ~ 20g/L and citric acid 0.001 ~ 5g/L;
Wherein, step D) in,
It is goose, wild goose, tortoise, ostrich or snake that described red blood cell is similar to the Magnocellular animal of human blood leukocyte;
The animal that described red blood cell is similar to human blood leukocyte's cellule is cat, dog, rabbit, monkey, ox, chicken or duck;
The animal that described red blood cell is similar to human blood Platelet Size is cat, horse, ox, sheep or camel.
2. the preparation method of Biologically-safe quality control material for blood tester according to claim 1, is characterized in that: steps A) in, the animal that described red blood cell is similar to human red cell size is cat, dog, pig, rabbit, monkey, ox, chicken or duck.
3. the preparation method of Biologically-safe quality control material for blood tester according to claim 1, is characterized in that: step e) in, the addition of formaldehyde is 40:1 ~ 1:1 with the erythrocytic volume ratio collected.
4. the preparation method of Biologically-safe quality control material for blood tester according to claim 1, is characterized in that: the microbiotic in described cell maintenance liquid is selected from penicillins, aminoglycoside and amide-type antibiotic any one or two or more combinations.
5. the preparation method of Biologically-safe quality control material for blood tester according to claim 1, is characterized in that: the polysaccharide in described cell maintenance liquid is selected from the combination of any one or more in sucrose, glucose, fructose, galactose, lactose, maltose, starch and dextrin.
6. according to the Quality Control thing that the preparation method according to any one of Claims 1 to 5 prepares.
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