CN103267838B - Quality control material and calibration material for verifying hematology analyzer and preparation method thereof - Google Patents
Quality control material and calibration material for verifying hematology analyzer and preparation method thereof Download PDFInfo
- Publication number
- CN103267838B CN103267838B CN 201310179485 CN201310179485A CN103267838B CN 103267838 B CN103267838 B CN 103267838B CN 201310179485 CN201310179485 CN 201310179485 CN 201310179485 A CN201310179485 A CN 201310179485A CN 103267838 B CN103267838 B CN 103267838B
- Authority
- CN
- China
- Prior art keywords
- cell
- blood
- supernatant
- treating fluid
- add
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Abstract
The invention discloses a quality control material and a calibration material for verifying hematology analyzer. Various liquids are integrated; a cell treating liquid integrates the functions of anti-coagulation, blood cell preservation and cell solidification stabilization; secondly, a manufacturing process is optimized, and pre-treatment is carried out immediately once blood is sampled, so that on one hand, the blood is prevented from coagulation and hemolysis, and on the other hand the immobilization of the blood cells is ultimately realized, and at the same time the activity of fresh blood is not lost; and by utilizing the process, not only the freshness of the blood cells is determined, but also the aging of the blood cells alive is determined, subsequently, by utilizing a centrifugation and separation technique, expected blood cells are extracted, and finally cell simulants of different concentrations are added so as to synthesize a complete standard substance; such process is low in production cost, the produced calibration material and the quality control material are high in yield; and the synthesized calibration material and the quality control material are stable in activity and longer in aging.
Description
[technical field]
The present invention relates to a kind of when verification cellanalyzer quality-control product used and calibration object.
[background technology]
The existing quality-control product of degree of accuracy, stability and accuracy and the making of calibration object that is used for verification cellanalyzer mainly relies on human blood and pig blood etc., but human blood or its goods are subject to country as commodity forbids the restriction of selling, and pig blood and human blood cell differ greatly, although cost is low, effect and human blood differ greatly.In addition, also there are some blood cell analysis instrument producers that supporting quality-control product and calibration object is provided, but theirs is expensive, and enjoy again the restriction of traffic condition and supply chain when import quality-control product and calibration object, so produce of home make cheap and good-quality quality-control product and calibration object seems too impatient to wait.
The applicant has proposed to replace human blood to manufacture the imagination of quality-control product and calibration object with monkey blood the earliest, and the application for a patent for invention that number of patent application is 201110000498.8 proposed in 2011, experimental study is for many years passed through in this invention, can successfully replace human blood for the manufacture of quality-control product and calibration object monkey blood.But it is complicated to some extent in technology, cause quality-control product or calibration object to be not easy to create, at recipe ingredient, use and show slightly dull in addition, such as adding after anti-coagulants in monkey whole blood blood plasma, though can guarantee that blood does not solidify, haemolysis not, but if this blood can not get timely processing, can there is haemolysis and then lose efficacy in it equally very soon; In addition, it is simple and coarse that manufacturing process also shows slightly, after anti-freezing, preservation, separating-purifying, interpolation cell analogue, just add stabilizing agent, such process can not guarantee cytoactive and integrality, cell may be before not adding stabilizing agent just not solidify or haemolysis causes blood cannot continue use again, so the calibration object for preparing of this process and the low qualified of quality-control product, thereby has greatly increased production cost, and activity stabilized poor, timeliness is partially short.
[summary of the invention]
The object of the invention is to overcome the deficiencies in the prior art, provide a kind of activity stabilized, timeliness is longer, has not only met national health requirement, but also the degree of accuracy of verification cellanalyzer and the quality-control product of stability more accurately.
Another object of the present invention is to overcome the deficiencies in the prior art, provides a kind of activity stabilized, and timeliness is longer, has not only met national health requirement, but also the degree of accuracy of verification cellanalyzer and the calibration object of stability more accurately.
Another object of the present invention has been to provide the preparation method for quality-control product and the calibration object of verification cellanalyzer.
In order to solve the technical matters of above-mentioned existence, the present invention adopts following technical proposals:
A kind of quality-control product for verification cellanalyzer, it includes monkey whole blood, cell treating fluid, Hydroxyethyl Starch 40 sodium chloride injections and cell analogue, wherein cell treating fluid divides to join for 4～6 times and in monkey whole blood, makes cell curing liquid, except the cell treating fluid adding for the last time, after adding cell treating fluid, fully mix, centrifugal and the supernatant of separating is all given up at every turn; The cell treating fluid adding first and the proportioning between monkey whole blood are 1:2.5～5, and the deal supernatant separated with the last time, that give up of the cell treating fluid backward at every turn adding equates; In gained cell curing liquid, press 1.5～6.5 * 10
12the unit capacity of/L is extracted monkey whole blood red blood cell and is gone out supernatant by centrifuging, adds Hydroxyethyl Starch 40 sodium chloride injections with the supernatant equal volume of just having given up after the supernatant of giving up 75% ~ 85% volume again, this:
The raw material of described cell treating fluid forms to be had: NaH2PO4,2-hydroxy propane-1,2,3-sodium tricarboxylate, 3-hydroxyl-3-carboxyl glutaric acid, NH4Cl, C6H12O6, C6H14O6, C5H5N5, C5H8O2, (CH2O) n, NaN3, sodium chloride and pure water, their proportioning is: 5～10g:3～6g:4～10g:1～3g:20～50g:10～50g:0.5～5g:0.05～1ml:0.05～1ml:0.5～1.5g:1～9g:1000ml;
The raw material of described cell analogue is the latex particle of mammiferous haemocyte or diameter 1～10 μ m.
In the improvement project to the above-mentioned quality-control product for verification cellanalyzer, the cell treating fluid and the proportioning between monkey whole blood that add are first 1:5; Described NaH2PO4,2-hydroxy propane-1,2,3-sodium tricarboxylate, 3-hydroxyl-3-carboxyl glutaric acid, NH4Cl, C6H12O6, C6H14O6, C5H5N5, C5H8O2, (CH2O) n, NaN3, the proportioning of sodium chloride and pure water is 6g:5.25g:5.5g:2g:30g:20g:1g:0.1ml:0.1ml:0.5g:5g:1000ml.
In the improvement project to the above-mentioned quality-control product for verification cellanalyzer, the cell treating fluid and the proportioning between monkey whole blood that add are first 1:4; Described NaH2PO4,2-hydroxy propane-1,2,3-sodium tricarboxylate, 3-hydroxyl-3-carboxyl glutaric acid, NH4Cl, C6H12O6, C6H14O6, C5H5N5, C5H8O2, (CH2O) n, NaN3, the proportioning of sodium chloride and pure water is 6g:5.25g:5.5g:2g:30g:20g:1g:0.05ml:0.05ml:0.5g:6g:1000ml.
A preparation method for the quality-control product of verification cellanalyzer, it includes following steps:
1), prepare cell treating fluid: in the ratio of proportioning: 5～10g:3～6g:4～10g:1～3g:20～50g:10～50g:0.5～5g:0.05～1ml:0.05～1ml:0.5～1.5g:1～9g:1000ml, take respectively or measure NaH2PO4,2-hydroxy propane-1,2,3-sodium tricarboxylate, 3-hydroxyl-3-carboxyl glutaric acid, NH4Cl, C6H12O6, C6H14O6, C5H5N5, C5H8O2, (CH2O) n, NaN3, sodium chloride and pure water, then mixing and stirring, after filtration, standby after sterilizing;
2), blood sampling pre-service: adopt monkey blood to the glass container of having sterilized, the ratio by cell treating fluid with 1:2.5～5 adds in its monkey blood immediately, fully mixes and seals, standing 1 ~ 3 hour;
3), cell curing is processed: get above-mentioned monkey blood pre-service product, fully mix, centrifugal, separation obtains supernatant first, it is all extracted out and is given up, again add inward the cell treating fluid with the supernatant equal volume of just having given up, fully mix, centrifugal, separation obtains secondary supernatant, also it is all extracted out and is given up, the cell treating fluid that the supernatant volume that repeats to give up with a upper procedure 2～4 times add with this equates, and mix, centrifugal, extract out and give up whole supernatants, the cell treating fluid of the supernatant equal volume that is with gives up for the last time finally adding, fully mix, the standing monkey blood consolidation liquid that becomes,
4), filtration: adopt blood transfusion apparatus to filter impurity and the grumeleuse in monkey blood consolidation liquid, the blood leaching after fully mixing flows into a bottle end by wall, to prevent haemolysis destruction cell, detects subsequently cell content and other parameter;
5), separation and Extraction: get the blood having filtered, by 1.5～6.5 * 10
12the unit capacity of/L is extracted required monkey whole blood red blood cell; Again mix and centrifuging goes out supernatant, extract and give up the supernatant of 75%～85% volume out, then add Hydroxyethyl Starch 40 sodium chloride injections with the supernatant equal volume of just having given up, fully mix, standing;
6), add cell analogue: add the latex particle of mammiferous haemocyte or diameter 1～10 μ m as cell analogue, when it is used for substituting leucocyte, by 2～25 * 10
9the unit capacity of/L is added, when being used for replacement blood platelet, by 30～900 * 10
9the unit capacity of/L is added;
7), test assignment: randomly draw some blood that is divided in test tube, on corresponding cellanalyzer, test on request or repeatedly detect the content of various cells and calculate assignment uncertainty by grain count instrument.
In the improvement project to the preparation method of the above-mentioned quality-control product for verification cellanalyzer, in described step 2, the ratio by cell treating fluid with 1:5 adds wherein immediately, fully mixes and seals, standing 2 hours; In described step 3, repeating to add the number of times with the equiponderant cell treating fluid of a upper procedure is 3 times; In described step 5, extract and give up the supernatant of 80% volume out, then add Hydroxyethyl Starch 40 sodium chloride injections with the supernatant equal volume of just having given up.
In the improvement project to the preparation method of the above-mentioned quality-control product for verification cellanalyzer, in described step 2, the ratio by cell treating fluid with 1:4 adds wherein immediately, fully mixes and seals, standing 3 hours; In described step 3, repeating to add the number of times with the equiponderant cell treating fluid of a upper procedure is 3 times; In described step 5, extract and give up the supernatant of 80% volume out, then add Hydroxyethyl Starch 40 sodium chloride injections with the supernatant equal volume of just having given up.
A kind of calibration object for verification cellanalyzer, it includes monkey whole blood, cell treating fluid, Hydroxyethyl Starch 40 sodium chloride injections and cell analogue, wherein cell treating fluid divides to join for 4 ~ 6 times and in monkey whole blood, makes cell curing liquid, and the cell treating fluid adding first and the proportioning between monkey whole blood are 1:2.5～5, the deal of the cell treating fluid backward at every turn adding supernatant separated with the last time, that give up equates; In gained cell curing liquid, press 1.5 ~ 6.5 * 10
12the unit capacity of/L is extracted monkey whole blood red blood cell and is gone out supernatant by centrifuging, adds Hydroxyethyl Starch 40 sodium chloride injections with the supernatant equal volume of just having given up after the supernatant of giving up 75% ~ 85% volume again, this:
The raw material of described cell treating fluid forms to be had: NaH2PO4,2-hydroxy propane-1,2,3-sodium tricarboxylate, 3-hydroxyl-3-carboxyl glutaric acid, NH4Cl, C6H12O6, C6H14O6, C5H5N5, C5H8O2, (CH2O) n, NaN3, sodium chloride and pure water, their proportioning is: 5～10g:3～6g:4～10g:1～3g:20～50g:10～50g:0.5～5g:0.05～1ml:0.05～1ml:0.5～1.5g:1～9g:1000ml;
The raw material of described cell analogue is the latex particle of diameter 1～10 μ m.
In the improvement project to the above-mentioned calibration object for verification cellanalyzer, the cell treating fluid and the proportioning between monkey whole blood that add are first 1:5; Described NaH2PO4,2-hydroxy propane-1,2,3-sodium tricarboxylate, 3-hydroxyl-3-carboxyl glutaric acid, NH4Cl, C6H12O6, C6H14O6, C5H5N5, C5H8O2, (CH2O) n, NaN3, the proportioning of sodium chloride and pure water is 6g:5.25g:5.5g:2g:30g:20g:1g:0.1ml:0.1ml:0.5g:5g:1000ml.
In the improvement project to the above-mentioned calibration object for verification cellanalyzer, the cell treating fluid and the proportioning between monkey whole blood that add are first 1:4; Described NaH2PO4,2-hydroxy propane-1,2,3-sodium tricarboxylate, 3-hydroxyl-3-carboxyl glutaric acid, NH4Cl, C6H12O6, C6H14O6, C5H5N5, C5H8O2, (CH2O) n, NaN3, the proportioning of sodium chloride and pure water is 6g:5.25g:5.5g:2g:30g:20g:1g:0.05ml:0.05ml:0.5g:6g:1000ml.
A preparation method for the calibration object of verification cellanalyzer, it includes following steps:
1), prepare cell treating fluid: in the ratio of proportioning: 5～10g:3～6g:4～10g:1～3g:20～50g:10～50g:0.5～5g:0.05～1ml:0.05～1ml:0.5～1.5g:1～9g:1000ml, take respectively or measure NaH2PO4,2-hydroxy propane-1,2,3-sodium tricarboxylate, 3-hydroxyl-3-carboxyl glutaric acid, NH4Cl, C6H12O6, C6H14O6, C5H5N5, C5H8O2, (CH2O) n, NaN3, sodium chloride and pure water, then mixing and stirring, after filtration, standby after sterilizing;
2), blood sampling pre-service: adopt monkey blood to the glass container of having sterilized, the ratio by cell treating fluid with 1:2.5～5 adds in its monkey blood immediately, fully mixes and seals, standing 1～3 hour;
3), cell curing is processed: get above-mentioned monkey blood pre-service product, fully mix, centrifugal, separation obtains supernatant first, it is all extracted out and is given up, again add inward the cell treating fluid with the supernatant equal volume of just having given up, fully mix, centrifugal, separation obtains secondary supernatant, also it is all extracted out and is given up, the cell treating fluid that the supernatant volume that repeats to give up with a upper procedure 2～4 times add with this equates, and mix, centrifugal, extract out and give up whole supernatants, the cell treating fluid of the supernatant equal volume that is with gives up for the last time finally adding, fully mix, the standing monkey blood consolidation liquid that becomes,
4), filtration: adopt blood transfusion apparatus to filter impurity and the grumeleuse in monkey blood consolidation liquid, the blood leaching after fully mixing flows into a bottle end by wall, to prevent haemolysis destruction cell, detects subsequently cell content and other parameter;
5), separation and Extraction: get the blood having filtered, by 1.5～6.5 * 10
12the unit capacity of/L is extracted required monkey whole blood red blood cell; Again mix and centrifuging goes out supernatant, extract and give up the supernatant of 75%～85% volume out, then add Hydroxyethyl Starch 40 sodium chloride injections with the supernatant equal volume of just having given up, fully mix, standing;
6), add cell analogue: add the latex particle of diameter 1～10 μ m as cell analogue, when it is used for substituting leucocyte, by 2～25 * 10
9the unit capacity of/L is added, when being used for replacement blood platelet, by 30～900 * 10
9the unit capacity of/L is added;
7), test assignment: randomly draw some blood that is divided in test tube, on corresponding cellanalyzer, test on request or repeatedly detect the content of various cells and calculate assignment uncertainty by grain count instrument.
In the improvement project to the preparation method of the above-mentioned calibration object for verification cellanalyzer, in described step 2, the ratio by cell treating fluid with 1:5 adds wherein immediately, fully mixes and seals, standing 2 hours; In described step 3, repeating to add the number of times with the equiponderant cell treating fluid of a upper procedure is 3 times; In described step 5, extract and give up the supernatant of 80% volume out, then add Hydroxyethyl Starch 40 sodium chloride injections with the supernatant equal volume of just having given up.
In the improvement project to the preparation method of the above-mentioned calibration object for verification cellanalyzer, in described step 2, the ratio by cell treating fluid with 1:4 adds wherein immediately, fully mixes and seals, standing 3 hours; In described step 3, repeating to add the number of times with the equiponderant cell treating fluid of a upper procedure is 3 times; In described step 5, extract and give up the supernatant of 80% volume out, then add Hydroxyethyl Starch 40 sodium chloride injections with the supernatant equal volume of just having given up.
Compared with prior art, the invention has the beneficial effects as follows: due to various liquid is integrated, cell treating fluid has merged anti-freezing, preserved haemocyte and the stable effect of cell curing, its suboptimization manufacturing process, after blood sampling, carry out at once pre-service, one guarantees that blood does not solidify not haemolysis, two carry out stabilized cell environment, keep the fresh activity of blood, add again cell treating fluid rinsing and curing haemocyte repeatedly, finally realize solidifying of haemocyte, do not lose it as the activity performance of new blood simultaneously, this road technique not only determines the fresh and alive degree of haemocyte, also determining the timeliness of haemocyte survival, then by centrifugal and isolation technics, extract needed haemocyte, the cell analogue that finally adds variable concentrations, thereby synthetic complete standard substance, the production cost of this technological process is low, and the calibration object of producing and the yield rate of quality-control product are high, make the activity stabilized of the quality-control product that is synthesized and calibration object, timeliness is longer, when testing by cellanalyzer or artificial counting tests, find that it has identical effect with human blood quality-control product and calibration object.
Below in conjunction with embodiment, the present invention is described in further detail:
[embodiment]
Defect and the deficiency of the technical scheme that is 201110000498.8 for number of patent application aspect two of recipe ingredient and manufacturing process, the present invention has carried out many experiments again, first improved recipe ingredient, and various liquid is integrated, cell treating fluid has merged anti-freezing, has preserved haemocyte and the stable effect of cell curing.Its suboptimization manufacturing process, after blood sampling, carry out at once pre-service, one guarantees that blood does not solidify not haemolysis, two carry out stabilized cell environment, keep the fresh activity of blood, add again cell treating fluid rinsing and curing haemocyte repeatedly, finally realize solidifying of haemocyte, do not lose it as the activity performance of new blood simultaneously, this road technique not only determines the fresh and alive degree of haemocyte, also determining the timeliness of haemocyte survival, then by centrifugal and isolation technics, extract needed haemocyte, the cell analogue that finally adds variable concentrations, thereby synthetic complete quality-control product and calibration object, the production cost of this technological process is low, and the quality-control product of producing and the yield rate of calibration object are high, the quality-control product being synthesized and calibration object activity stabilized, timeliness is longer, when testing by cellanalyzer or artificial counting tests, find that it has identical effect with human blood quality-control product and calibration object.
In the present invention, quality-control product for verification cellanalyzer, by monkey whole blood, cell treating fluid, Hydroxyethyl Starch 40 sodium chloride injections and cell analogue form, monkey whole blood is through the pre-service of cell treating fluid, prevent haemocyte haemolysis and aggegation, guarantee the fresh of blood, haemocyte is complete and active, again by the curing processing of cell treating fluid, the living environment of stabilised blood cell, increase the stability of haemocyte film, slow down the spending rate of haemocyte energy, extend the holding time of haemocyte, form stable haemocyte, and add therein Hydroxyethyl Starch 40 sodium chloride injections of certain volume, the last cell analogue of variable concentrations that adds as requested again, a thereby synthetic quality-control product that can substitute human blood.
This quality-control product, include monkey whole blood, cell treating fluid, Hydroxyethyl Starch 40 sodium chloride injections and cell analogue, wherein cell treating fluid divides to join for 4 ~ 6 times and in monkey whole blood, makes cell curing liquid, except last, after adding cell treating fluid, fully mix at every turn, centrifugal and the supernatant of separating is all given up, and the cell treating fluid adding first and the proportioning between monkey whole blood are 1:2.5～5, the deal (referring to volume at this) of the cell treating fluid backward at every turn adding supernatant separated with the last time, that give up equates; In gained cell curing liquid, press 1.5～6.5 * 10
12the unit capacity of/L is extracted monkey whole blood red blood cell and is gone out supernatant by centrifuging, adds Hydroxyethyl Starch 40 sodium chloride injections with the supernatant equal volume of just having given up after the supernatant of giving up 75%～85% volume again, this:
The raw material of described cell treating fluid forms to be had: NaH2PO4,2-hydroxy propane-1,2,3-sodium tricarboxylate, 3-hydroxyl-3-carboxyl glutaric acid, NH4Cl, C6H12O6, C6H14O6, C5H5N5, C5H8O2, (CH2O) n, NaN3, sodium chloride and pure water, their proportioning is: 5～10g:3～6g:4～10g:1～3g:20～50g:10～50g:0.5～5g:0.05～1ml:0.05～1ml:0.5～1.5g:1～9g:1000ml.Wherein, the effect of cell treating fluid prevents monkey whole blood coagulation, for haemocyte provides nutrition, keeps haemocyte active, and haemocyte is carried out pre-service and solidify processing, and extends the cell survival life-span; With 2-hydroxy propane-1,2,3-sodium tricarboxylate and 3-hydroxyl-3-carboxyl glutaric acid are as anti-coagulants, and the buffering that they form, to regulating and stabilizing solution acidity-basicity ph, maintains blood acid-base balance; NaH2PO4 prevents erythrocyte aggregation, for energy metabolism of erythrocyte provides phosphate, slows down 2,3-diphosphoglyceric acid decline rate; NH4Cl can reduce carbon dioxide content in blood plasma; C6H12O6 is the necessary nutritional labeling of erythrocyte metabolism, can extend the erythrocytic holding time, and prevents haemolysis, and can make the organophosphorus in cell disappear slowly, prevents the damage that red blood cell stores; C6H14O6 cell membrane has protective effect, and does not affect the function of cell membrane, thereby has improved erythrocytic motility rate again; C5H5N5 can promote red blood cell ATP synthetic, extends erythrocytic storage life, and strengthens red blood cell and put oxygen function; C5H8O2 and (CH2O) n can solidify cell; NaN3 can be anticorrosion; Sodium chloride is used for the osmotic pressure of regulator solution.
The raw material of described cell analogue is the latex particle of mammiferous haemocyte or diameter 1～10 μ m, and it can substitute the large cellule of ad eundem, when the cell analogue adding is used for substituting leucocyte, by 2～25 * 10
9the unit capacity of/L is added, when the cell analogue adding is used for replacement blood platelet, by 30～900 * 10
9the unit capacity of/L is added.Because latex particle is stable, can guarantee the accuracy of quantity, and coordinate the different fixing histogram of the required making of different instruments or scatter diagram for data analysis.
The main production flow process of this quality-control product is as follows:
Prepare cell treating fluid-blood sampling pre-service-cell curing-filtration-separation and Extraction-interpolation cell analogue-assignment.Wherein:
1), prepare cell treating fluid: in the ratio of proportioning: 5～10g:3～6g:4～10g:1～3g:20～50g:10～50g:0.5～5g:0.05～1ml:0.05～1ml:0.5～1.5g:1～9g:1000ml, take respectively or measure NaH2PO4,2-hydroxy propane-1,2,3-sodium tricarboxylate, 3-hydroxyl-3-carboxyl glutaric acid, NH4Cl, C6H12O6, C6H14O6, C5H5N5, C5H8O2, (CH2O) n, NaN3, sodium chloride and pure water, then mixing and stirring, after filtration, standby after sterilizing;
2), blood sampling pre-service: adopt monkey blood to the glass container of having sterilized, the ratio by cell treating fluid with 1:2.5～5 adds in its monkey blood immediately, fully mixes and seals, standing 1 ~ 3 hour;
3), cell curing is processed: get above-mentioned monkey blood pre-service product, fully mix, centrifugal, separation obtains supernatant first, it is all extracted out and is given up, again add inward the cell treating fluid with the supernatant equal volume of just having given up, fully mix, centrifugal, separation obtains secondary supernatant, also it is all extracted out and is given up, the cell treating fluid that the supernatant volume that repeats to give up with a upper procedure 2～4 times add with this equates, and mix, centrifugal, extract out and give up whole supernatants, the cell treating fluid of the supernatant equal volume that is with gives up for the last time finally adding, fully mix, the standing monkey blood consolidation liquid that becomes,
4), filtration: adopt blood transfusion apparatus to filter impurity and the grumeleuse in monkey blood consolidation liquid, the blood leaching after fully mixing flows into a bottle end by wall, to prevent haemolysis destruction cell, detects subsequently cell content and other parameter;
5), separation and Extraction: according to the quantity of the fixed centrifugal number of times of the height of cell numerical value and extraction cell, get the blood having filtered and extract required monkey whole blood red blood cell, and press 1.5～6.5 * 10
12the unit capacity of/L is extracted; Again mix and centrifuging goes out supernatant, extract and give up the supernatant of 75%～85% volume out, then add Hydroxyethyl Starch 40 sodium chloride injections with the supernatant equal volume of just having given up, fully mix, standing;
6), add cell analogue: the raw material of the cell analogue wherein adding is the latex particle of mammiferous haemocyte or diameter 1～10 μ m, when it is used for substituting leucocyte, by 2～25 * 10
9the unit capacity of/L is added, when being used for replacement blood platelet, by 30～900 * 10
9the unit capacity of/L is added;
7), test assignment: randomly draw some blood that is divided in test tube, on corresponding cellanalyzer, test on request or repeatedly detect the content of various cells and calculate assignment uncertainty by grain count instrument.
Because calibration object of the present invention is compared with quality-control product, their difference is just only selected the latex particle of diameter 1～10 μ m at the cell analogue of calibration object.
With the preparation of quality-control product, give an example below.
Embodiment mono-, and the preparation flow of quality-control product is:
1), prepare cell treating fluid: by weight or volume get respectively the NaH2PO4 of 5g, 2-hydroxy propane-1 of 3g, 2,3-sodium tricarboxylate, 3-hydroxyl-3-carboxyl glutaric acid of 4g, the NH4Cl of 1g, the C6H12O6 of 20g, the C6H14O6 of 10g, the C5H5N5 of 0.5g, the C5H8O2 of 0.05ml, (CH2O) n of 0.05ml, the NaN3 of 0.5g, the sodium chloride of 9g and the pure water of 1000ml, then mixing and stirring, after filtration, standby after sterilizing;
2), blood sampling pre-service: the monkey blood of adopting 1000ml is to the glass container of having sterilized, by 200ml(in the ratio of 1:5) cell treating fluid add immediately wherein, fully mix and seal, standing 1 hour;
3), cell curing is processed: get above-mentioned monkey blood pre-service product, fully mix, centrifugal, separation obtains supernatant first, it is all extracted out and is given up, again add inward the cell treating fluid with the supernatant equal volume of just having given up, fully mix, centrifugal, separation obtains secondary supernatant, also it is all extracted out and is given up, the cell treating fluid that the supernatant volume that repeats to give up with a upper procedure 2 times add with this equates, and mix, centrifugal, extract out and give up whole supernatants, the cell treating fluid of the supernatant equal volume that is with gives up for the last time finally adding, fully mix, the standing monkey blood consolidation liquid that becomes,
4), filtration: adopt blood transfusion apparatus to filter impurity and the grumeleuse in monkey blood consolidation liquid, the blood leaching after fully mixing flows into a bottle end by wall, to prevent haemolysis destruction cell, detects subsequently cell content and other parameter;
5), separation and Extraction: decide the quantity of centrifugal number of times and extraction cell according to the height of cell numerical value, get the blood having filtered, by 1.5 * 10
12the unit capacity of/L is extracted monkey whole blood red blood cell, again mix also centrifuging and go out supernatant (about 1000ml), extract and give up the supernatant (750ml) of 75% volume out, Hydroxyethyl Starch 40 sodium chloride injections of the supernatant equal volume that then adds and just given up (being 750ml), fully mix, standing;
6), add cell analogue: add the haemocyte of cat blood, when cat blood is used for substituting leucocyte, by 2 * 10
9the unit capacity of/L is added, when being used for replacement blood platelet, by 30 * 10
9the unit capacity of/L is added;
7), test assignment: randomly draw some blood that is divided in test tube, on corresponding cellanalyzer, test on request or repeatedly detect the content of various cells and calculate assignment uncertainty by grain count instrument.
Embodiment bis-, and the preparation flow of quality-control product is:
1), prepare cell treating fluid: by weight or volume get respectively the NaH2PO4 of 10g, 2-hydroxy propane-1 of 6g, 2,3-sodium tricarboxylate, 3-hydroxyl-3-carboxyl glutaric acid of 10g, the NH4Cl of 3g, the C6H12O6 of 50g, the C6H14O6 of 50g, the C5H5N5 of 5g, the C5H8O2 of 1ml, (CH2O) n of 1ml, the NaN3 of 1.5g, the sodium chloride of 1g and the pure water of 1000ml, then mixing and stirring, after filtration, standby after sterilizing;
2), blood sampling pre-service: the monkey blood of adopting 1000ml is to the glass container of having sterilized, by 400ml(in the ratio of 1:2.5) cell treating fluid add immediately wherein, fully mix and seal, standing 3 hours;
3), cell curing is processed: get above-mentioned monkey blood pre-service product, fully mix, centrifugal, separation obtains supernatant first, it is all extracted out and is given up, again add inward the cell treating fluid with the supernatant equal volume of just having given up, fully mix, centrifugal, separation obtains secondary supernatant, also it is all extracted out and is given up, the cell treating fluid that the supernatant volume that repeats to give up with a upper procedure 4 times add with this equates, and mix, centrifugal, extract out and give up whole supernatants, the cell treating fluid of the supernatant equal volume that is with gives up for the last time finally adding, fully mix, the standing monkey blood consolidation liquid that becomes,
4), filtration: adopt blood transfusion apparatus to filter impurity and the grumeleuse in monkey blood consolidation liquid, the blood leaching after fully mixing flows into a bottle end by wall, to prevent haemolysis destruction cell, detects subsequently cell content and other parameter;
5), separation and Extraction: decide the quantity of centrifugal number of times and extraction cell according to the height of cell numerical value, get the blood having filtered, by 6.5 * 10
12the unit capacity of/L is extracted monkey whole blood red blood cell, again mix also centrifuging and go out supernatant (about 800ml), extract and give up the supernatant (680ml) of 85% volume out, Hydroxyethyl Starch 40 sodium chloride injections of the supernatant equal volume that then adds and just given up (being 680ml), fully mix, standing;
6), add cell analogue: adding diameter is that the latex particle of 10 μ m substitutes leucocyte, unit capacity by 25 * 109/L is added cell analogue, and when being used for replacement blood platelet, adding diameter is that the latex particle of 1.5 μ m substitutes, and concentration is added by the unit capacity of 900 * 109/L;
7), test assignment: randomly draw some blood that is divided in test tube, on corresponding cellanalyzer, test on request or repeatedly detect the content of various cells and calculate assignment uncertainty by grain count instrument.
Embodiment tri-, and the preparation flow of quality-control product is:
1), prepare cell treating fluid: by weight or volume get respectively the NaH2PO4 of 7.5g, 2-hydroxy propane-1 of 4.5g, 2,3-sodium tricarboxylate, 3-hydroxyl-3-carboxyl glutaric acid of 7g, the NH4Cl of 2g, the C6H12O6 of 35g, the C6H14O6 of 30g, the C5H5N5 of 2.75g, the C5H8O2 of 0.52ml, (CH2O) n of 0.52ml, the NaN3 of 1g, the sodium chloride of 5g and the pure water of 1000ml, then mixing and stirring, after filtration, standby after sterilizing;
2), blood sampling pre-service: the monkey blood of adopting 750ml is to the glass container of having sterilized, by 200ml(in the ratio of 1:3.75) cell treating fluid add immediately wherein, fully mix and seal, standing 2.5 hours;
3), cell curing is processed: get above-mentioned monkey blood pre-service product, fully mix, centrifugal, separation obtains supernatant first, it is all extracted out and is given up, again add inward the cell treating fluid with the supernatant equal volume of just having given up, fully mix, centrifugal, separation obtains secondary supernatant, also it is all extracted out and is given up, the cell treating fluid that the supernatant volume that repeats to give up with a upper procedure 3 times add with this equates, and mix, centrifugal, extract out and give up whole supernatants, the cell treating fluid of the supernatant equal volume that is with gives up for the last time finally adding, fully mix, the standing monkey blood consolidation liquid that becomes,
4), filtration: adopt blood transfusion apparatus to filter impurity and the grumeleuse in monkey blood consolidation liquid, the blood leaching after fully mixing flows into a bottle end by wall, to prevent haemolysis destruction cell, detects subsequently cell content and other parameter;
5), separation and Extraction: decide the quantity of centrifugal number of times and extraction cell according to the height of cell numerical value, get the blood having filtered, by 4.0 * 10
12the unit capacity of/L is extracted monkey whole blood red blood cell, again mix also centrifuging and go out supernatant (about 600ml), extract and give up the supernatant (468ml) of 78% volume out, Hydroxyethyl Starch 40 sodium chloride injections of the supernatant equal volume that then adds and just given up (being 468ml), fully mix, standing;
6), add cell analogue: adding diameter is that the latex particle of 6 μ m substitutes leucocyte, unit capacity by 13.5 * 109/L is added cell analogue, and when being used for replacement blood platelet, adding diameter is that the latex particle of 3 μ m substitutes, and concentration is added by the unit capacity of 465 * 109/L;
7), test assignment: randomly draw some blood that is divided in test tube, on corresponding cellanalyzer, test on request or repeatedly detect the content of various cells and calculate assignment uncertainty by grain count instrument.
Embodiment tetra-, and the preparation flow of quality-control product is:
1), prepare cell treating fluid: by weight or volume get respectively the NaH2PO4 of 6g, 2-hydroxy propane-1 of 5.25g, 2,3-sodium tricarboxylate, 3-hydroxyl-3-carboxyl glutaric acid of 5.5g, the NH4Cl of 2g, the C6H12O6 of 30g, the C6H14O6 of 20g, the C5H5N5 of 1g, the C5H8O2 of 0.1ml, (CH2O) n of 0.1ml, the NaN3 of 0.5g, the sodium chloride of 5g and the pure water of 1000ml, then mixing and stirring, after filtration, standby after sterilizing;
2), blood sampling pre-service: the monkey blood of adopting 500ml is to the glass container of having sterilized, by 100ml(in the ratio of 1:5) cell treating fluid add immediately wherein, fully mix and seal, standing 2 hours;
3), cell curing is processed: get above-mentioned monkey blood pre-service product, fully mix, centrifugal, separation obtains supernatant first, it is all extracted out and is given up, again add inward the cell treating fluid with the supernatant equal volume of just having given up, fully mix, centrifugal, separation obtains secondary supernatant, also it is all extracted out and is given up, the cell treating fluid that the supernatant volume that repeats to give up with a upper procedure 3 times add with this equates, and mix, centrifugal, extract out and give up whole supernatants, the cell treating fluid of the supernatant equal volume that is with gives up for the last time finally adding, fully mix, the standing monkey blood consolidation liquid that becomes,
4), filtration: adopt blood transfusion apparatus to filter impurity and the grumeleuse in monkey blood consolidation liquid, the blood leaching after fully mixing flows into a bottle end by wall, to prevent haemolysis destruction cell, detects subsequently cell content and other parameter;
5), separation and Extraction: decide the quantity of centrifugal number of times and extraction cell according to the height of cell numerical value, get the blood having filtered, by 2.3 * 10
12the unit capacity of/L is extracted monkey whole blood red blood cell, again mix also centrifuging and go out supernatant (about 400ml), extract and give up the supernatant (320ml) of 80% volume out, Hydroxyethyl Starch 40 sodium chloride injections of the supernatant equal volume that then adds and just given up (being 320ml), fully mix, standing;
6), add cell analogue: adding diameter is that the latex particle of 6 μ m substitutes leucocyte, unit capacity by 7.5 * 109/L is added cell analogue, and when being used for replacement blood platelet, adding diameter is that the latex particle of 2.5 μ m substitutes, and concentration is added by the unit capacity of 250 * 109/L;
7), test assignment: randomly draw some blood that is divided in test tube, on corresponding cellanalyzer, test on request or repeatedly detect the content of various cells and calculate assignment uncertainty by grain count instrument.
Embodiment five, and the preparation flow of quality-control product is:
1), prepare cell treating fluid: by weight or volume get respectively the NaH2PO4 of 6g, 2-hydroxy propane-1 of 5.25g, 2,3-sodium tricarboxylate, 3-hydroxyl-3-carboxyl glutaric acid of 5.5g, the NH4Cl of 2g, the C6H12O6 of 30g, the C6H14O6 of 20g, the C5H5N5 of 1g, the C5H8O2 of 0.05ml, (CH2O) n of 0.05ml, the NaN3 of 0.5g, the sodium chloride of 6g and the pure water of 1000ml, then mixing and stirring, after filtration, standby after sterilizing;
2), blood sampling pre-service: the monkey blood of adopting 500ml is to the glass container of having sterilized, by 125ml(in the ratio of 1:4) cell treating fluid add immediately wherein, fully mix and seal, standing 3 hours;
3), cell curing is processed: get above-mentioned monkey blood pre-service product, fully mix, centrifugal, separation obtains supernatant first, it is all extracted out and is given up, again add inward the cell treating fluid with the supernatant equal volume of just having given up, fully mix, centrifugal, separation obtains secondary supernatant, also it is all extracted out and is given up, the cell treating fluid that the supernatant volume that repeats to give up with a upper procedure 3 times add with this equates, and mix, centrifugal, extract out and give up whole supernatants, the cell treating fluid of the supernatant equal volume that is with gives up for the last time finally adding, fully mix, the standing monkey blood consolidation liquid that becomes,
4), filtration: adopt blood transfusion apparatus to filter impurity and the grumeleuse in monkey blood consolidation liquid, the blood leaching after fully mixing flows into a bottle end by wall, to prevent haemolysis destruction cell, detects subsequently cell content and other parameter;
5), separation and Extraction: decide the quantity of centrifugal number of times and extraction cell according to the height of cell numerical value, get the blood having filtered, by 5.3 * 10
12the unit capacity of/L is extracted monkey whole blood red blood cell, again mix also centrifuging and go out supernatant (about 390ml), extract and give up the supernatant (312ml) of 80% volume out, Hydroxyethyl Starch 40 sodium chloride injections of the supernatant equal volume that then adds and just given up (being 312ml), fully mix, standing;
6), add cell analogue: adding diameter is that the latex particle of 6 μ m substitutes leucocyte, unit capacity by 19 * 109/L is added cell analogue, and when being used for replacement blood platelet, adding diameter is that the latex particle of 4 μ m substitutes, and concentration is added by the unit capacity of 695 * 109/L;
7), test assignment: randomly draw some blood that is divided in test tube, on corresponding cellanalyzer, test on request or repeatedly detect the content of various cells and calculate assignment uncertainty by grain count instrument.
Embodiment six, and the preparation flow of quality-control product is:
1), prepare cell treating fluid: by weight or volume get respectively the NaH2PO4 of 10g, 2-hydroxy propane-1 of 4g, 2,3-sodium tricarboxylate, 3-hydroxyl-3-carboxyl glutaric acid of 5g, the NH4Cl of 2g, the C6H12O6 of 40g, the C6H14O6 of 30g, the C5H5N5 of 2g, the C5H8O2 of 0.5ml, (CH2O) n of 0.5ml, the NaN3 of 1g, the sodium chloride of 6g and the pure water of 1000ml, then mixing and stirring, after filtration, standby after sterilizing;
2), blood sampling pre-service: the monkey blood of adopting 600ml is to the glass container of having sterilized, by 200ml(in the ratio of 1:3) cell treating fluid add immediately wherein, fully mix and seal, standing 3 hours;
3), cell curing is processed: get above-mentioned monkey blood pre-service product, fully mix, centrifugal, separation obtains supernatant first, it is all extracted out and is given up, again add inward the cell treating fluid with the supernatant equal volume of just having given up, fully mix, centrifugal, separation obtains secondary supernatant, also it is all extracted out and is given up, the cell treating fluid that the supernatant volume that repeats to give up with a upper procedure 2 times add with this equates, and mix, centrifugal, extract out and give up whole supernatants, the cell treating fluid of the supernatant equal volume that is with gives up for the last time finally adding, fully mix, the standing monkey blood consolidation liquid that becomes,
4), filtration: adopt blood transfusion apparatus to filter impurity and the grumeleuse in monkey blood consolidation liquid, the blood leaching after fully mixing flows into a bottle end by wall, to prevent haemolysis destruction cell, detects subsequently cell content and other parameter;
5), separation and Extraction: decide the quantity of centrifugal number of times and extraction cell according to the height of cell numerical value, get the blood having filtered, by 4.8 * 10
12the unit capacity of/L is extracted monkey whole blood red blood cell, again mix also centrifuging and go out supernatant (about 550ml), extract and give up the supernatant (451ml) of 82% volume out, Hydroxyethyl Starch 40 sodium chloride injections of the supernatant equal volume that then adds and just given up (being 451ml), fully mix, standing;
6), add cell analogue: add the haemocyte of sheep blood, when it is used for substituting leucocyte, by 12.5 * 10
9the unit capacity of/L is added, when being used for replacement blood platelet, by 500 * 10
9/ L unit capacity is added;
7), test assignment: randomly draw some blood that is divided in test tube, on corresponding cellanalyzer, test on request or repeatedly detect the content of various cells and calculate assignment uncertainty by grain count instrument.
WBC(white blood cell count(WBC) from haemocyte), RBC(red blood cell count(RBC)), HGB(haemoglobin), PLT(platelet count), the MCV(mean corpuscular volume) coefficient of variation (the Coefficient of Variance of these five parameters, be abbreviated as CV%), the cell survival term of validity evaluates stability and the cell survival of quality-control product or calibration object, thereby finds out implementation result:
The parameters of quality-control product
| ? | WBC | RBC | HGB | PLT | MCV | The uncork term of validity | The sealing term of validity |
| Embodiment 1 | 2.5% | 2.0% | 5.0% | 8.0% | 5.0% | 15 days | 85 days |
| Embodiment 2 | 3.0% | 2.5% | 6.0% | 9.0% | 6.0% | 15 days | 80 days |
| Embodiment 3 | 2.2% | 1.8% | 4.5% | 7.5% | 4.5% | 15 days | 95 days |
| Embodiment 4 | 1.0% | 0.6% | 2.0% | 3.5% | 2.5% | 20 days | 105 days |
| Embodiment 5 | 1.2% | 0.8% | 2.0% | 3.5% | 2.8% | 20 days | 100 days |
| Embodiment 6 | 1.8% | 1.0% | 2.0% | 4.0% | 3.0% | 15 days | 90 days |
The parameters of calibration object
| ? | WBC | RBC | HGB | PLT | MCV | The uncork term of validity | The sealing term of validity |
| Embodiment 1 | 2.5% | 2.0% | 5.0% | 8.0% | 5.0% | 8 days | 85 days |
| Embodiment 2 | 3.0% | 2.5% | 6.0% | 9.0% | 6.0% | 8 days | 80 days |
| Embodiment 3 | 2.2% | 1.8% | 4.5% | 7.5% | 4.5% | 8 days | 95 days |
| Embodiment 4 | 1.0% | 0.6% | 2.0% | 3.5% | 2.5% | 12 days | 105 days |
| Embodiment 5 | 1.2% | 0.8% | 2.0% | 3.5% | 2.8% | 12 days | 100 days |
| Embodiment 6 | 1.8% | 1.0% | 2.0% | 4.0% | 3.0% | 8 days | 90 days |
Can obtain in sum: 1, the present invention simplifies the constituent of quality-control product and calibration object, anti-coagulants, preservation liquid and stabilizing agent are integrated into cell treating fluid, increase and decrease to some extent on recipe ingredient simultaneously, cell treating fluid after integration not only comprises the effect of anti-freezing, preservation and stabilized cell, also in whole technological process, cytoactive maintenance and cells survival is played to decisive role; 2, optimized technological process, new blood has been carried out to pre-service at once, stablized its fresh and alive degree, then cell curing has been processed, kept cytoactive, extended the expiration date; Once 3, this project for the production of, can greatly reduce equally the production cost of like product, alleviate better the situation of domestic quality-control product and calibration object market supply and demand anxiety.
Claims (12)
1. the quality-control product for verification cellanalyzer, it is characterized in that: include monkey whole blood, cell treating fluid, Hydroxyethyl Starch 40 sodium chloride injections and cell analogue, wherein cell treating fluid divides to join for 4～6 times and in monkey whole blood, makes cell curing liquid, except the cell treating fluid adding for the last time, after adding cell treating fluid, fully mix, centrifugal and the supernatant of separating is all given up at every turn; The cell treating fluid adding first and the volume proportion between monkey whole blood are 1:2.5～5, and the deal supernatant separated with the last time, that give up of the cell treating fluid backward at every turn adding equates; In gained cell curing liquid, press 1.5～6.5 * 10
12the unit capacity of/L is extracted monkey whole blood red blood cell and is gone out supernatant by centrifuging, after the supernatant of giving up 75% ~ 85% volume, add again Hydroxyethyl Starch 40 sodium chloride injections with the supernatant equal volume of just having given up, next add again cell analogue, this:
The raw material of described cell treating fluid forms to be had: NaH
2pO
4, 2-hydroxy propane-1,2,3-sodium tricarboxylate, 3-hydroxyl-3-carboxyl glutaric acid, NH
4cl, C
6h
12o
6, C
6h
14o
6, C
5h
5n
5, C
5h
8o
2, (CH
2o)
n, NaN
3, sodium chloride and pure water, their proportioning is: 5～10g:3～6g:4～10g:1～3g:20～50g:10～50g:0.5～5g:0.05～1mL:0.05～1 mL:0.5～1.5g:1～9g:1000 mL;
The raw material of described cell analogue is the latex particle of mammiferous haemocyte or diameter 1～10 μ m; When the cell analogue adding is used for substituting leucocyte, by 2～25 * 10
9the unit capacity of/L is added, when the cell analogue adding is used for replacement blood platelet, by 30～900 * 10
9the unit capacity of/L is added.
2. the quality-control product for verification cellanalyzer according to claim 1, is characterized in that: the cell treating fluid and the volume proportion between monkey whole blood that add are first 1:5;
Described NaH
2pO
4, 2-hydroxy propane-1,2,3-sodium tricarboxylate, 3-hydroxyl-3-carboxyl glutaric acid, NH
4cl, C
6h
12o
6, C
6h
14o
6,c
5h
5n
5, C
5h
8o
2, (CH
2o)
n, NaN
3, the proportioning of sodium chloride and pure water is 6g:5.25g:5.5g:2g:30g:20g:1g:0.1 mL:0.1 mL:0.5g:5g:1000 mL.
3. the quality-control product for verification cellanalyzer according to claim 1, is characterized in that: the cell treating fluid and the volume proportion between monkey whole blood that add are first 1:4;
Described NaH
2pO
4, 2-hydroxy propane-1,2,3-sodium tricarboxylate, 3-hydroxyl-3-carboxyl glutaric acid, NH
4cl, C
6h
12o
6, C
6h
14o
6, C
5h
5n
5, C
5h
8o
2, (CH
2o)
n, NaN
3, the proportioning of sodium chloride and pure water is 6g:5.25g:5.5g:2g:30g:20g:1g:0.05 mL:0.05 mL:0.5g:6g:1000 mL.
4. for a preparation method for the quality-control product of verification cellanalyzer, it includes following steps:
1), prepare cell treating fluid: in proportioning: the ratio of mL:0.05～1,5～10g:3～6g:4～10g:1～3g:20～50g:10～50g:0.5～5g:0.05～1 mL:0.5～1.5g:1～9g:1000 mL takes respectively or measure NaH
2pO
4, 2-hydroxy propane-1,2,3-sodium tricarboxylate, 3-hydroxyl-3-carboxyl glutaric acid, NH
4cl, C
6h
12o
6, C
6h
14o
6, C
5h
5n
5, C
5h
8o
2, (CH
2o)
n, NaN
3, sodium chloride and pure water, mixing and stirring then, after filtration, standby after sterilizing;
2), blood sampling pre-service: adopt monkey blood to the glass container of having sterilized, the volume ratio by cell treating fluid with 1:2.5～5 adds in its monkey blood immediately, fully mixes and seals, standing 1 ~ 3 hour;
3), cell curing is processed: get above-mentioned monkey blood pre-service product, fully mix, centrifugal, separation obtains supernatant first, it is all extracted out and is given up, again add inward the cell treating fluid with the supernatant equal volume of just having given up, fully mix, centrifugal, separation obtains secondary supernatant, also it is all extracted out and is given up, the cell treating fluid that the supernatant volume that repeats to give up with a upper procedure 2～4 times add with this equates, and mix, centrifugal, extract out and give up whole supernatants, the cell treating fluid of the supernatant equal volume that is with gives up for the last time finally adding, fully mix, the standing monkey blood consolidation liquid that becomes,
4), filtration: adopt blood transfusion apparatus to filter impurity and the grumeleuse in monkey blood consolidation liquid, the blood leaching after fully mixing flows into a bottle end by wall, to prevent haemolysis destruction cell, detects subsequently cell content and other parameter;
5), separation and Extraction: get the blood having filtered, by 1.5～6.5 * 10
12the unit capacity of/L is extracted required monkey whole blood red blood cell; Again mix and centrifuging goes out supernatant, extract and give up the supernatant of 75%～85% volume out, then add Hydroxyethyl Starch 40 sodium chloride injections with the supernatant equal volume of just having given up, fully mix, standing;
6), add cell analogue: add the latex particle of mammiferous haemocyte or diameter 1～10 μ m as cell analogue, when it is used for substituting leucocyte, by 2～25 * 10
9the unit capacity of/L is added, when being used for replacement blood platelet, by 30～900 * 10
9the unit capacity of/L is added;
7), test assignment: randomly draw some blood that is divided in test tube, on corresponding cellanalyzer, test on request or repeatedly detect the content of various cells and calculate assignment uncertainty by grain count instrument.
5. the preparation method of the quality-control product for verification cellanalyzer according to claim 4, is characterized in that:
In described step 2, the volume ratio by cell treating fluid with 1:5 adds wherein immediately, fully mixes and seals, standing 2 hours;
In described step 3, repeating to add the number of times with the equiponderant cell treating fluid of a upper procedure is 3 times;
In described step 5, extract and give up the supernatant of 80% volume out, then add Hydroxyethyl Starch 40 sodium chloride injections with the supernatant equal volume of just having given up.
6. the preparation method of the quality-control product for verification cellanalyzer according to claim 4, is characterized in that:
In described step 2, the volume ratio by cell treating fluid with 1:4 adds wherein immediately, fully mixes and seals, standing 3 hours;
In described step 3, repeating to add the number of times with the equiponderant cell treating fluid of a upper procedure is 3 times;
In described step 5, extract and give up the supernatant of 80% volume out, then add Hydroxyethyl Starch 40 sodium chloride injections with the supernatant equal volume of just having given up.
7. the calibration object for verification cellanalyzer, it is characterized in that: include monkey whole blood, cell treating fluid, Hydroxyethyl Starch 40 sodium chloride injections and cell analogue, wherein cell treating fluid divides to join for 4 ~ 6 times and in monkey whole blood, makes cell curing liquid, and the cell treating fluid adding first and the volume proportion between monkey whole blood are 1:2.5～5, the deal of the cell treating fluid backward at every turn adding supernatant separated with the last time, that give up equates; In gained cell curing liquid, press 1.5 ~ 6.5 * 10
12the unit capacity of/L is extracted monkey whole blood red blood cell and is gone out supernatant by centrifuging, after the supernatant of giving up 75% ~ 85% volume, add again Hydroxyethyl Starch 40 sodium chloride injections with the supernatant equal volume of just having given up, next add again cell analogue, this:
The raw material of described cell treating fluid forms to be had: NaH
2pO
4, 2-hydroxy propane-1,2,3-sodium tricarboxylate, 3-hydroxyl-3-carboxyl glutaric acid, NH
4cl, C
6h
12o
6, C
6h
14o
6, C
5h
5n
5, C
5h
8o
2, (CH
2o)
n, NaN
3, sodium chloride and pure water, their proportioning is: mL:0.05～1,5～10g:3～6g:4～10g:1～3g:20～50g:10～50g:0.5～5g:0.05～1 mL:0.5～1.5g:1～9g:1000 mL;
The raw material of described cell analogue is the latex particle of diameter 1～10 μ m, when it is used for substituting leucocyte, by 2～25 * 10
9the unit capacity of/L is added, when being used for replacement blood platelet, by 30～900 * 10
9the unit capacity of/L is added.
8. the calibration object for verification cellanalyzer according to claim 7, is characterized in that: the cell treating fluid and the volume proportion between monkey whole blood that add are first 1:5;
Described NaH
2pO
4, 2-hydroxy propane-1,2,3-sodium tricarboxylate, 3-hydroxyl-3-carboxyl glutaric acid, NH
4cl, C
6h
12o
6, C
6h
14o
6, C
5h
5n
5, C
5h
8o
2, (CH
2o)
n, NaN
3, the proportioning of sodium chloride and pure water is 6g:5.25g:5.5g:2g:30g:20g:1g:0.1 mL:0.1 mL:0.5g:5g:1000 mL.
9. the calibration object for verification cellanalyzer according to claim 7, is characterized in that: the cell treating fluid and the volume proportion between monkey whole blood that add are first 1:4;
Described NaH
2pO
4, 2-hydroxy propane-1,2,3-sodium tricarboxylate, 3-hydroxyl-3-carboxyl glutaric acid, NH
4cl, C
6h
12o
6, C
6h
14o
6, C
5h
5n
5, C
5h
8o
2, (CH
2o)
n, NaN
3, the proportioning of sodium chloride and pure water is 6g:5.25g:5.5g:2g:30g:20g:1g:0.05 mL:0.05 mL:0.5g:6g:1000 mL.
10. for a preparation method for the calibration object of verification cellanalyzer, it includes following steps:
1), prepare cell treating fluid: in proportioning: the ratio of mL:0.05～1,5～10g:3～6g:4～10g:1～3g:20～50g:10～50g:0.5～5g:0.05～1 mL:0.5～1.5g:1～9g:1000 mL takes respectively or measure NaH
2pO
4, 2-hydroxy propane-1,2,3-sodium tricarboxylate, 3-hydroxyl-3-carboxyl glutaric acid, NH
4cl, C
6h
12o
6, C
6h
14o
6, C
5h
5n
5, C
5h
8o
2, (CH
2o)
n, NaN
3, sodium chloride and pure water, mixing and stirring then, after filtration, standby after sterilizing;
2), blood sampling pre-service: adopt monkey blood to the glass container of having sterilized, the volume ratio by cell treating fluid with 1:2.5～5 adds in its monkey blood immediately, fully mixes and seals, standing 1～3 hour;
3), cell curing is processed: get above-mentioned monkey blood pre-service product, fully mix, centrifugal, separation obtains supernatant first, it is all extracted out and is given up, again add inward the cell treating fluid with the supernatant equal volume of just having given up, fully mix, centrifugal, separation obtains secondary supernatant, also it is all extracted out and is given up, the cell treating fluid that the supernatant volume that repeats to give up with a upper procedure 2～4 times add with this equates, and mix, centrifugal, extract out and give up whole supernatants, the cell treating fluid of the supernatant equal volume that is with gives up for the last time finally adding, fully mix, the standing monkey blood consolidation liquid that becomes,
4), filtration: adopt blood transfusion apparatus to filter impurity and the grumeleuse in monkey blood consolidation liquid, the blood leaching after fully mixing flows into a bottle end by wall, to prevent haemolysis destruction cell, detects subsequently cell content and other parameter;
5), separation and Extraction: get the blood having filtered, by 1.5～6.5 * 10
12the unit capacity of/L is extracted required monkey whole blood red blood cell; Again mix and centrifuging goes out supernatant, extract and give up the supernatant of 75%～85% volume out, then add Hydroxyethyl Starch 40 sodium chloride injections with the supernatant equal volume of just having given up, fully mix, standing;
6), add cell analogue: add the latex particle of diameter 1～10 μ m as cell analogue, when it is used for substituting leucocyte, by 2～25 * 10
9the unit capacity of/L is added, when being used for replacement blood platelet, by 30～900 * 10
9the unit capacity of/L is added;
7), test assignment: randomly draw some blood that is divided in test tube, on corresponding cellanalyzer, test on request or repeatedly detect the content of various cells and calculate assignment uncertainty by grain count instrument.
The preparation method of 11. calibration objects for verification cellanalyzer according to claim 10, is characterized in that:
In described step 2, the volume ratio by cell treating fluid with 1:5 adds wherein immediately, fully mixes and seals, standing 2 hours;
In described step 3, repeating to add the number of times with the equiponderant cell treating fluid of a upper procedure is 3 times;
In described step 5, extract and give up the supernatant of 80% volume out, then add Hydroxyethyl Starch 40 sodium chloride injections with the supernatant equal volume of just having given up.
The preparation method of 12. calibration objects for verification cellanalyzer according to claim 10, is characterized in that:
In described step 2, the volume ratio by cell treating fluid with 1:4 adds wherein immediately, fully mixes and seals, standing 3 hours;
In described step 3, repeating to add the number of times with the equiponderant cell treating fluid of a upper procedure is 3 times;
In described step 5, extract and give up the supernatant of 80% volume out, then add Hydroxyethyl Starch 40 sodium chloride injections with the supernatant equal volume of just having given up.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201310179485 CN103267838B (en) | 2013-05-15 | 2013-05-15 | Quality control material and calibration material for verifying hematology analyzer and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201310179485 CN103267838B (en) | 2013-05-15 | 2013-05-15 | Quality control material and calibration material for verifying hematology analyzer and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103267838A CN103267838A (en) | 2013-08-28 |
| CN103267838B true CN103267838B (en) | 2013-08-28 |
Family
ID=49011477
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201310179485 Active CN103267838B (en) | 2013-05-15 | 2013-05-15 | Quality control material and calibration material for verifying hematology analyzer and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103267838B (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105651568A (en) * | 2015-12-28 | 2016-06-08 | 西安交通大学第附属医院 | Preparation method of control material for blood cell analyzer |
| CN109470533B (en) * | 2018-10-24 | 2021-09-21 | 北京市临床检验中心 | Preparation method of human whole blood matrix quality control product for portable glucometer |
| CN112805567A (en) * | 2018-12-25 | 2021-05-14 | 深圳迈瑞生物医疗电子股份有限公司 | Erythrocyte-simulating particle, preparation method thereof and quality control substance or calibrator containing erythrocyte-simulating particle |
| CN111307695B (en) * | 2020-03-11 | 2022-06-14 | 深圳大学 | Calibration device and method of laser scattering classification device for cell counting |
| CN113959807A (en) * | 2021-10-26 | 2022-01-21 | 上海瀚诺威生物科技有限公司 | Preparation method of glycosylated hemoglobin calibration quality control product |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6448085B1 (en) * | 2001-04-13 | 2002-09-10 | Sysmex Corporation | Quality control material and calibrator for nucleated red blood cell tested on hematology analyzer |
| CN1873413A (en) * | 2006-06-27 | 2006-12-06 | 四川省迈克科技有限责任公司 | Quality control objects of whole blood, and production method |
| CN101672853A (en) * | 2009-09-28 | 2010-03-17 | 江西特康科技有限公司 | Blood cell analyzer calibrator and preparation process thereof |
| CN102435724A (en) * | 2011-01-04 | 2012-05-02 | 中山市滔略生物科技有限公司 | Quality control material and calibrator for calibrating blood cell analyzers and preparation method thereof |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1263266A (en) * | 1999-02-08 | 2000-08-16 | 刘剑雄 | Development of whole blood quality control substance in cell three-classification of hematology |
-
2013
- 2013-05-15 CN CN 201310179485 patent/CN103267838B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6448085B1 (en) * | 2001-04-13 | 2002-09-10 | Sysmex Corporation | Quality control material and calibrator for nucleated red blood cell tested on hematology analyzer |
| CN1873413A (en) * | 2006-06-27 | 2006-12-06 | 四川省迈克科技有限责任公司 | Quality control objects of whole blood, and production method |
| CN101672853A (en) * | 2009-09-28 | 2010-03-17 | 江西特康科技有限公司 | Blood cell analyzer calibrator and preparation process thereof |
| CN102435724A (en) * | 2011-01-04 | 2012-05-02 | 中山市滔略生物科技有限公司 | Quality control material and calibrator for calibrating blood cell analyzers and preparation method thereof |
Non-Patent Citations (2)
| Title |
|---|
| 寿好长.血细胞分析仪全血质控品的研制.《实用医技杂志》.2003,第10卷(第12期),第1332－1333页. |
| 血细胞分析仪全血质控品的研制;寿好长;《实用医技杂志》;20031231;第10卷(第12期);第1332－1333页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103267838A (en) | 2013-08-28 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103267838B (en) | Quality control material and calibration material for verifying hematology analyzer and preparation method thereof | |
| CN105246467B (en) | For stablizing the method for being encapsulated with the suspension of red blood cell of active constituent, suspension obtained | |
| JP4712135B2 (en) | Anticoagulation and storage solutions in the preparation of concentrated erythrocytes. | |
| CN102435724A (en) | Quality control material and calibrator for calibrating blood cell analyzers and preparation method thereof | |
| CN101672853B (en) | Blood cell analyzer calibrator and preparation process thereof | |
| CN101311724A (en) | Single group leucocyte emulation ion, calibrator comprising same and its preparation method | |
| CN105717312B (en) | A kind of red blood cell simulation particle, preparation method and Quality Control object or caliberator containing the simulation particle | |
| CN105454220B (en) | Placenta preservation method, placenta preservation solution and preparation method thereof | |
| JPWO2019230972A1 (en) | Composition, cell preservation composition, cell culture composition, cell preparation, method for producing an object containing microbubbles, method for storing cells, method for culturing cells, and method for producing a cell preparation. | |
| CN101181302A (en) | Upkeep liquid as well as preservative fluid for plastocyte | |
| US20160252434A1 (en) | Gamma sterilized dextran solutions and methods of use | |
| EP2839741A1 (en) | Apheresis platelets with fixed residual plasma volume | |
| CN109238927B (en) | Whole blood quality control substance and preparation method thereof | |
| Armenia et al. | Evaluation of Storage Length to Blood Component Platelet Concentrate Quality in the Blood Bank, Dr. Soetomo General Hospital, Surabaya, Indonesia. | |
| Gyongyossy‐Issa et al. | Novel system for storage of buffy‐coat‐derived platelet concentrates in a glucose‐based platelet additive solution: parameters and metabolism during storage and comparison to plasma | |
| CN102620950B (en) | Platelet preserving agent | |
| CN105746711A (en) | Milk product for improving sleep and preparation method thereof | |
| CN1256587C (en) | All blood quality controlling arlicles for blood rheology and their preparation | |
| Izadpanahi et al. | Evaluation of biochemical parameters of platelet concentrates stored in plasma or in a platelet additive solution (Composol) | |
| CN110997933B (en) | Methods and compositions for stabilizing cell-free nucleic acids and cells | |
| AU2021101429A4 (en) | Method for extracting flavonoids as active substances from honey | |
| CN114053422A (en) | Sodium carboxymethyl starch and its injection | |
| CN205460112U (en) | Separator is gathered to full -automatic whole blood | |
| CN107080970B (en) | A kind of subcritical dialysis process of water soluble ingredient | |
| CN106119183A (en) | Hyclone for Cell culture invitro and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |