CN115669648A - Erythrocyte preservation reagent and application thereof - Google Patents
Erythrocyte preservation reagent and application thereof Download PDFInfo
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Abstract
The invention relates to an erythrocyte preservation reagent and application thereof, belonging to the technical field of biology. The invention provides a red blood cell preservation reagent, which comprises the components of a solvent, glutaraldehyde and paraformaldehyde; the mass fraction of the glutaraldehyde in the solvent is 0.004-0.030%; the mass fraction of the paraformaldehyde in the solvent is 0.04-0.14%. The erythrocyte preservation reagent can form a cross-linked structure with protein on the surface of erythrocytes, so that stable 'carbon-nitrogen' covalent bonds are formed between the erythrocytes and are in close contact with the erythrocytes to fix erythrocyte membrane antigens, so that the volume of the erythrocytes is increased, the deformability is weakened, the osmotic fragility is weakened, the osmotic pressure resistance is improved, and the antigens are stable; research shows that when the erythrocyte preservation reagent is used for preserving erythrocytes at 4 ℃, the preservation period can exceed 6 months when the mass fraction of the erythrocytes is 5 percent, and the erythrocyte preservation reagent is not easy to hemolysis and has good antigen stability.
Description
Technical Field
The invention relates to an erythrocyte preservation reagent and application thereof, belonging to the technical field of biology.
Background
The erythrocyte reagent has wide application range and is mainly used for blood type reverse typing detection, blood type antibody titer determination and erythrocyte antibody screening and identification. The great britain pharmacopoeia particularly emphasizes two aspects of detecting surface antigen of red blood cells and antibody in blood plasma before blood transfusion, and the next operation can be carried out only if the positive and negative typing is consistent; blood group antibody titer determination and erythrocyte antibody screening and identification are the most basic guarantees of human blood product quality, the blood product must detect ABO blood group antibody titer to effectively avoid hemolytic transfusion reaction, and leakage detection of other irregular antibodies can cause delayed hemolytic transfusion reaction.
However, at the present stage, the storage life of commercial erythrocyte reagents used for blood type reverse typing detection, blood type antibody titer determination and erythrocyte antibody screening and identification is only 2-3 months under the condition of matching with low temperature (generally 4 ℃), and the commercial erythrocyte reagents are easy to dissolve in blood, have low antigen stability and seriously affect clinical application. Compared with the erythrocyte reagent, the preservation period of the erythrocyte by using ultra-low temperature freezing and freeze-drying is longer, generally more than 6 months, and the erythrocyte is not easy to be hemolyzed and has better antigen stability. However, ultra-low temperature freezing (generally-80 ℃) requires an ultra-low temperature refrigerator or a liquid nitrogen tank, consumes a large amount of liquid nitrogen or electric energy, is difficult to transport, has a complex operation process of rewarming and washing to remove the cryoprotective agent (generally high-concentration glycerol), wastes time and labor, and is difficult to ensure that the cryoprotective agent is added and removed at a reasonable cost in a reasonable time on the premise of not losing excessive red blood cells; the freeze-drying requires special freeze-drying equipment and freeze-drying protective agent, and has the disadvantages of complex operation process, time consumption and labor consumption.
Disclosure of Invention
In order to solve the problems, the invention provides an erythrocyte preservation reagent, and the components of the erythrocyte preservation reagent comprise a solvent, glutaraldehyde and paraformaldehyde; the mass fraction of the glutaraldehyde in the solvent is 0.004-0.006%; the mass fraction of the paraformaldehyde in the solvent is 0.04-0.06%.
In one embodiment of the invention, the mass fraction of glutaraldehyde in the solvent is 0.005%; the mass fraction of the paraformaldehyde in the solvent is 0.05%.
In one embodiment of the invention, the solvent is a solution containing trisodium citrate, potassium dihydrogen phosphate, disodium hydrogen phosphate dodecahydrate, sodium chloride, glucose, sucrose, adenine, creatinine, kanamycin, chloramphenicol, glutaraldehyde and/or paraformaldehyde.
The invention also provides a method for preserving red blood cells, which comprises the following steps: erythrocytes were added to the above-mentioned erythrocyte preservation reagent to preserve erythrocytes.
In one embodiment of the present invention, the method is: and adding the red blood cells into the red blood cell preservation reagent until the mass fraction of the red blood cells is 1-10% so as to preserve the red blood cells.
In one embodiment of the present invention, the method is: the red blood cells are added to the red blood cell preservation reagent to be 5% in mass fraction so as to be preserved.
In one embodiment of the present invention, the method is: diluting the red blood cells with a solvent until the mass fraction of the red blood cells is 1-20% to obtain a red blood cell diluent; mixing the erythrocyte diluent with the erythrocyte preservation reagent according to a volume ratio of 1: 0.5-2, rolling for 5-20 min by a roller at 18-25 ℃ by a roller-mixing instrument, and then taking packed red blood cells; the packed red blood cells are diluted to 1 to 10 mass percent by using the red blood cell preservation reagent to preserve the red blood cells.
In one embodiment of the present invention, the method is: diluting the red blood cells with a solvent until the mass fraction of the red blood cells is 10% to obtain a red blood cell diluent; mixing the erythrocyte diluent and the erythrocyte preservation reagent according to the volume ratio of 1:1, after mixing, rolling for 10min by a roller at the temperature of 18-25 ℃ by a roller-mixing instrument, and then taking packed red blood cells; the packed erythrocytes were diluted to 5% by mass with the above-mentioned erythrocyte preservation reagent to preserve erythrocytes.
The invention also provides the application of the erythrocyte preservation reagent or the erythrocyte preservation method in preserving the erythrocytes.
The invention also provides a red blood cell preservation solution, which is obtained by preserving red blood cells by using the method for preserving red blood cells; the components of the erythrocyte preservation solution comprise a solvent, glutaraldehyde, paraformaldehyde and erythrocytes; the mass fraction of the glutaraldehyde in the solvent is 0.004-0.006%; the mass fraction of the paraformaldehyde in the solvent is 0.04-0.06%; the mass fraction of the red blood cells in the solvent is 1-10%.
In one embodiment of the invention, the mass fraction of glutaraldehyde in the solvent is 0.005%; the mass fraction of the paraformaldehyde in the solvent is 0.05 percent; the mass fraction of the red blood cells in the solvent is 5%.
The invention also provides the application of the erythrocyte preservation reagent or the erythrocyte preservation method or the erythrocyte preservation liquid in blood group reverse typing detection, blood group antibody titer determination or erythrocyte antibody screening and identification, and the application has the non-disease diagnosis and treatment purposes.
The technical scheme of the invention has the following advantages:
the invention provides a red blood cell preservation reagent, which comprises the components of a solvent, glutaraldehyde and paraformaldehyde; the mass fraction of the glutaraldehyde in the solvent is 0.004-0.006%; the mass fraction of the paraformaldehyde in the solvent is 0.04-0.06%. The erythrocyte preservation reagent can fix erythrocyte membrane antigens, so that the volume of erythrocytes is increased, the deformability is weakened, the osmotic fragility is weakened, the osmotic pressure resistance is improved, and the antigens are stable; research shows that the erythrocyte preservation reagent is used for preserving erythrocytes at 4 ℃, when the mass fraction of the erythrocytes is 5%, the preservation period can reach 6 months, hemolysis is not easy to occur, and the antigen stability is good, so the erythrocyte preservation solution obtained by preserving the erythrocytes by using the erythrocyte preservation reagent can be used as the erythrocyte reagent and is widely applied to blood type reverse typing detection, blood type antibody titer determination or erythrocyte antibody screening and identification.
Further, the mass fraction of the glutaraldehyde in the solvent is 0.005%; the mass fraction of the paraformaldehyde in the solvent is 0.05 percent; the erythrocyte preservation reagent under the formula has the best effect of preserving erythrocytes.
Drawings
FIG. 1: results of antigen intensity measurements (test tube method) of erythrocytes after storage in different erythrocyte storage reagents.
FIG. 2: results of antigen intensity measurements of erythrocytes after storage in different erythrocyte storage reagents (column gel method + use of antibody reagent). In FIG. 2, -A represents the result obtained by using an anti-A agent; -B represents the results obtained with the anti-B agent.
FIG. 3: results of antigen intensity measurements of red blood cells after storage in different red blood cell storage reagents (column gel method + use of plasma samples). In FIG. 3, A represents the results obtained using type A plasma; b represents the results obtained with type B plasma; o represents the result obtained by using type O plasma; AB represents the results obtained using type AB plasma.
FIG. 4: the results of the measurement of free hemoglobin in the supernatant (type A erythrocytes) were obtained after the erythrocytes had been stored in the reagent for preservation of erythrocytes for various times.
FIG. 5: the results of the measurement of the free hemoglobin of the supernatant (B-type erythrocytes) were obtained after the erythrocytes had been stored in the reagent for preserving erythrocytes for various times.
FIG. 6: layering of supernatant and hematocrit after preservation of erythrocytes in different erythrocyte preservation reagents (24 h of preservation).
FIG. 7: stratification of supernatant and hematocrit after preservation of erythrocytes in different erythrocyte preservation reagents (6 month preservation).
FIG. 8: the dispersion of erythrocytes after storage in different erythrocyte preservation reagents.
Detailed Description
The following examples are provided to further understand the present invention, not to limit the scope of the present invention, but to provide the best mode, not to limit the content and the protection scope of the present invention, and any product similar or similar to the present invention, which is obtained by combining the present invention with other prior art features, falls within the protection scope of the present invention.
The following examples do not show specific experimental procedures or conditions, and can be performed according to the procedures or conditions of the conventional experimental procedures described in the literature in the field. The reagents or instruments used are not indicated by manufacturers, and are all conventional reagent products which can be obtained commercially.
Example 1: erythrocyte preservation reagent and preparation method thereof
The embodiment provides an erythrocyte preservation reagent, which comprises the components of normal saline, glutaraldehyde and paraformaldehyde; the mass fraction of the glutaraldehyde in the physiological saline is 0.005%; the mass fraction of the paraformaldehyde in the physiological saline is 0.05%.
The preparation method of the erythrocyte preservation reagent comprises the following steps:
diluting glutaraldehyde (purchased from Chinese medicinal company) with normal saline to the mass fraction of 0.01% to obtain glutaraldehyde diluent; diluting paraformaldehyde (purchased from Chinese medicinal company) with normal saline to a mass fraction of 0.1% to obtain paraformaldehyde diluent; mixing glutaraldehyde diluent and paraformaldehyde diluent according to a volume ratio of 1:1 to obtain a reagent for preserving erythrocytes 1 (preservation at 4 ℃).
Example 2: erythrocyte preservation reagent and preparation method thereof
The present example provides an erythrocyte preservation reagent based on example 1: adjusting the mass fraction of glutaraldehyde in physiological saline to 0.2%, and simultaneously adjusting the mass fraction of paraformaldehyde in physiological saline to 0%, so as to obtain a red blood cell preservation reagent 2;
adjusting the mass fraction of glutaraldehyde in the physiological saline to 0.18%, and simultaneously adjusting the mass fraction of paraformaldehyde in the physiological saline to 0%, so as to obtain a red blood cell preservation reagent 3;
adjusting the mass fraction of glutaraldehyde in the physiological saline to 0.16%, and simultaneously adjusting the mass fraction of paraformaldehyde in the physiological saline to 0% to obtain a red blood cell preservation reagent 4;
adjusting the mass fraction of glutaraldehyde in the physiological saline to 0.14%, and simultaneously adjusting the mass fraction of paraformaldehyde in the physiological saline to 0% to obtain a red blood cell preservation reagent 5;
adjusting the mass fraction of glutaraldehyde in the physiological saline to 0.12%, and simultaneously adjusting the mass fraction of paraformaldehyde in the physiological saline to 0%, so as to obtain a red blood cell preservation reagent 6;
adjusting the mass fraction of glutaraldehyde in physiological saline to 0.1%, and simultaneously adjusting the mass fraction of paraformaldehyde in physiological saline to 0%, so as to obtain a red blood cell preservation reagent 7;
adjusting the mass fraction of glutaraldehyde in the physiological saline to 0.04%, and simultaneously adjusting the mass fraction of paraformaldehyde in the physiological saline to 0% to obtain a red blood cell preservation reagent 8;
adjusting the mass fraction of glutaraldehyde in the physiological saline to 0.03%, and simultaneously adjusting the mass fraction of paraformaldehyde in the physiological saline to 0%, so as to obtain a red blood cell preservation reagent 9;
adjusting the mass fraction of glutaraldehyde in the physiological saline to 0.02%, and simultaneously adjusting the mass fraction of paraformaldehyde in the physiological saline to 0%, so as to obtain a red blood cell preservation reagent 10;
adjusting the mass fraction of glutaraldehyde in physiological saline to 0.01%, and simultaneously adjusting the mass fraction of paraformaldehyde in physiological saline to 0%, so as to obtain a red blood cell preservation reagent 11;
adjusting the mass fraction of glutaraldehyde in the physiological saline to 0.05%, and simultaneously adjusting the mass fraction of paraformaldehyde in the physiological saline to 0.2%, so as to obtain a red blood cell preservation reagent 12;
adjusting the mass fraction of glutaraldehyde in the physiological saline to 0.04%, and simultaneously adjusting the mass fraction of paraformaldehyde in the physiological saline to 0.2%, so as to obtain a red blood cell preservation reagent 13;
adjusting the mass fraction of glutaraldehyde in the physiological saline to 0.03%, and simultaneously adjusting the mass fraction of paraformaldehyde in the physiological saline to 0.2%, so as to obtain a red blood cell preservation reagent 14;
adjusting the mass fraction of glutaraldehyde in physiological saline to 0.02%, and simultaneously adjusting the mass fraction of paraformaldehyde in physiological saline to 0.2%, so as to obtain a red blood cell preservation reagent 15;
adjusting the mass fraction of glutaraldehyde in physiological saline to 0.01%, and simultaneously adjusting the mass fraction of paraformaldehyde in physiological saline to 0.2%, so as to obtain a red blood cell preservation reagent 16;
adjusting the mass fraction of glutaraldehyde in the physiological saline to 0.01%, and simultaneously adjusting the mass fraction of paraformaldehyde in the physiological saline to 0.05% to obtain an erythrocyte preservation reagent 17;
adjusting the mass fraction of glutaraldehyde in physiological saline to 0.02%, and simultaneously adjusting the mass fraction of paraformaldehyde in physiological saline to 0.1%, to obtain erythrocyte preservation reagent 18;
adjusting the mass fraction of glutaraldehyde in the physiological saline to 0.02%, and simultaneously adjusting the mass fraction of paraformaldehyde in the physiological saline to 0.05%, so as to obtain a red blood cell preservation reagent 19;
adjusting the mass fraction of glutaraldehyde in physiological saline to 0.005%, and simultaneously adjusting the mass fraction of paraformaldehyde in physiological saline to 0.1%, to obtain erythrocyte preservation reagent 20;
the mass fraction of glutaraldehyde in physiological saline was adjusted to 0.005%, and the mass fraction of paraformaldehyde in physiological saline was adjusted to 0.05%, to obtain erythrocyte preservation reagent 21.
Example 3: erythrocyte preservation solution and preparation method thereof
The embodiment provides a red blood cell preservation solution, which comprises the components of normal saline, glutaraldehyde, paraformaldehyde and red blood cells; the mass fraction of the glutaraldehyde in the solvent is 0.005%; the mass fraction of the paraformaldehyde in the solvent is 0.05 percent; the mass fraction of the red blood cells in the physiological saline is 5%.
The preparation method of the erythrocyte preservation solution comprises the following steps:
washing A, B, O, AB type fresh erythrocyte with physiological saline (centrifugation parameter of washing: 1200rpm for 5 min) for three times, and collecting packed erythrocyte A; diluting packed red blood cells A with normal saline until the mass fraction is 10% to obtain red blood cell diluent; the volume ratio of the red blood cell diluent to the red blood cell preservation reagent 1 in example 1 is 1:1, after mixing, firstly rolling for 10min by a roller at room temperature (25 ℃) by a roller and mixing instrument, then washing for four times by physiological saline (the centrifugal parameter of washing is 1200rpm and centrifugation is 5 min), and reserving packed red blood cells B; packed red blood cells B were diluted to 5% by mass with the red blood cell preservation reagent of example 1, to obtain a red blood cell preservation solution.
Example 4: erythrocyte preservation solution and preparation method thereof
The present embodiment provides a red blood cell preservation solution, which is prepared in example 3 by: erythrocyte preservation solutions were obtained by replacing the erythrocyte preservation reagent 1 of example 1 with the erythrocyte preservation reagent of example 2.
Experimental example 1: erythrocyte preservation reagent for checking preservation capability of erythrocyte
The experimental example verifies the preservation capability of the erythrocyte preservation reagent according to erythrocyte reagent related index detection, wherein the erythrocyte reagent related index detection comprises antigen intensity, antibody specificity, supernatant free hemoglobin, preservation appearance and dispersity. The specific verification process is as follows:
experiment one: antigen intensity detection
The antigen intensity of erythrocytes in the erythrocyte preservation solution obtained by the method of examples 3 to 4 was measured by the test tube method (see table 1 for the criterion of agglutination intensity by test tube method) and the column gel method (see table 2 for the criterion of agglutination intensity by column gel method);
wherein, the test tube method is as follows: taking non-hydroformylation A-type erythrocytes as a control, preparing hydroformylation A-type erythrocytes into erythrocyte suspension with the mass fraction of 5% by using erythrocyte preservation solution; placing the erythrocyte suspension at room temperature (25 deg.C) for different time (24 h, 1month, 2month, 3month, 4month, 5month, 6 month), mixing 100 μ L of erythrocyte suspension with 100 μ L of anti-A reagent (purchased from Shanghai blood organism), centrifuging at 1000rpm for 1min, and collecting precipitate; visually observing the agglutination strength of the precipitate, and comparing the result with the non-hydroformylation erythrocytes of the control group, wherein the detection and comparison results are shown in figure 1;
the column gel method comprises the following steps: taking non-hydroformylation A, B type red blood cells as a reference, preparing hydroformylation A, B type red blood cells into red blood cell suspensions with mass fractions of 0.8% by using red blood cell preservation solution; after the erythrocyte suspension was left at room temperature (25 ℃) for various times (1 month, 6 month), 50. Mu.L of the erythrocyte suspension was mixed with 50. Mu.L of an anti-A agent (purchased from Haematology Shanghai), an anti-B agent (purchased from Haematology Shanghai) and human A, B, O, AB type 5363 plasma (from Michidai Union Hospital, jilin university), centrifuged at 900rpm for 2min, and finally centrifuged at 1800rpm for 8min to obtain a precipitate; the distribution of the red blood cells in the microcolumn gel card is observed, and the result is compared with the non-hydroformylation A, B type red blood cells of the control group, and the detection and comparison results are shown in figures 2 to 3.
As shown in FIG. 1, the agglutination strength of erythrocytes in 0.005% glutaraldehyde and 0.05% paraformaldehyde erythrocyte preservative solution was 4+ within 6 months, which is consistent with the results for erythrocytes that were not aldehyde-treated; the agglutination strength of the erythrocytes in 0.01 percent of glutaraldehyde and 0.05 percent of paraformaldehyde erythrocyte preservation solution is more than 2+ within 3 months, and then the tendency of weakening is presented; the antigen intensity of erythrocytes in 0.02% glutaraldehyde and 0.05% paraformaldehyde erythrocyte preservation solution is 2+ in 24 hours, and the rest time is less than 2+.
As shown in fig. 2 to 3, after the red blood cells in the 0.005% glutaraldehyde and 0.05% paraformaldehyde red blood cell preservation solution reacted with the reagent or plasma after 6 months of storage, the background of the negative result in the microcolumn gel card was clear, while the background of the negative result in the microcolumn gel card was unclear after the red blood cells in the other red blood cell preservation solution reacted with the reagent or plasma.
Experiment two: antibody specificity detection
The antibody specificity of erythrocytes in the erythrocyte preservation solution obtained by the method of examples 3 to 4 was detected by a test tube method (see Table 1 for the criterion of agglutination strength by test tube method);
wherein, the test tube method is as follows: taking non-hydroformylation A, B type red blood cells as a reference, and preparing non-hydroformylation A, B type red blood cells into red blood cell suspensions with mass fractions of 5% by using red blood cell preservation solution; placing the erythrocyte suspension at room temperature (25 ℃) for 24 hours, mixing 100 mu L of the erythrocyte suspension with 100 mu L of an anti-A reagent (purchased from Shanghai blood biology), an anti-B reagent (purchased from Shanghai blood biology), an anti-AB reagent (purchased from Shanghai blood biology) and human AB type erythrocyte plasma (from Mitsui Union Hospital, jilin university), centrifuging at 1000rpm for 1min, and taking a precipitate; the agglutination intensity of the precipitates was visually observed, and the results were compared with control group of non-hydroformylated A, B type erythrocytes, and the results of detection and comparison are shown in table 3.
As shown in table 3, the specificity of the antibody was the same for the hydroformylated erythrocytes as for the non-hydroformylated erythrocytes; taking the hydroformylation type A erythrocytes and non-hydroformylation type A erythrocytes as examples, 4+ agglutination condition occurs between the hydroformylation type A erythrocytes and the anti-A antibody reagent and the anti-AB reagent; the agglutination result was negative by reaction with the anti-B antibody reagent.
Experiment three: supernatant free hemoglobin detection
Taking non-hydroformylation A, B type red blood cells as a reference, preparing non-hydroformylation A, B type red blood cells into red blood cell suspensions with mass fractions of 5% by taking red blood cell preservation solution; the erythrocyte suspension was left at room temperature (25 ℃) for different periods of time (1 month, 2month, 3month, 4month, 5month, 6 month), and then the supernatant free hemoglobin in the erythrocyte suspension was detected using Nanjing-constructed hemoglobin detection kit (colorimetry), and the detection results are shown in FIGS. 4 to 5.
Comparing the ratio of the free hemoglobin of the supernatant of the non-hydroformylated erythrocyte and the hydroformylation erythrocyte at the same time, as shown in 4-5, the ratio of the free hemoglobin of the supernatant obtained by hemolysis of the non-hydroformylated erythrocyte in the first two months is less than the ratio of the free hemoglobin of the supernatant obtained by hemolysis of the erythrocyte in 0.005% glutaraldehyde and 0.05% erythrocyte preservative solution; the free hemoglobin of the supernatant of the non-hydroformylation red blood cells is obviously increased after 4 months, and is more than 0.005 percent of glutaraldehyde and 0.05 percent of the free hemoglobin of the supernatant obtained by hemolysis of the red blood cells in the red blood cell preservation solution.
Experiment four: appearance detection
Taking non-hydroformylation A, B type red blood cells as a reference, preparing hydroformylation A, B type red blood cells into red blood cell suspensions with mass fractions of 5% by taking red blood cell preservation solution; the erythrocyte suspension is placed at room temperature (25 ℃) for different time (24 h, 6 month), the layering condition of the supernatant and the hematocrit in the erythrocyte preservation solution in different preservation periods is observed by naked eyes, and the observation result is shown in figures 6-7.
Fig. 6 shows that the upper layer liquid is clear, and the upper layer liquid is obviously layered with red blood cells. FIG. 7 shows that the upper liquid layer of the non-hydroformylated red blood cells turns red, the upper liquid layer is obviously layered with the red blood cells, and the hydroformylation red blood cells still keep the upper liquid layer clear, and the upper liquid layer is obviously layered with the red blood cells.
Experiment five: dispersion detection
Taking non-hydroformylation A, B type red blood cells and fresh A, B type red blood cells as controls, preparing hydroformylation A, B type red blood cells into red blood cell suspensions with mass fractions of 5% by using red blood cell preservation solution; the dispersion of erythrocytes in the erythrocyte preservation solution was observed under a mirror after the erythrocyte suspension was left at room temperature (25 ℃) for 6month, and the results of the observation are shown in FIG. 8.
As shown in FIG. 8, the hydroformylation erythrocytes showed good dispersibility under the lens and no aggregation or stacking at all when stored for 6 months.
TABLE 1 test tube method agglutination strength interpretation standards
TABLE 2 determination of agglutination strength by column gel method
| Strength of agglutination | Interpretation standards |
| ++++ | The red blood cell complex is located on the surface of the gel |
| +++ | Most of the red blood cell complex is on the surface of the gel, and a small part is on the middle upper part of the gel |
| ++ | Most of the compound is located in the middle of the glue, and a small part of the compound is located in the middle upper part of the glue |
| + | The compound is located near the bottom of the glue |
| - | Depositing red blood cells at the bottom of the tip of the gel tube of the microcolumn after centrifugation |
TABLE 3 antibody specificity test results for erythrocytes in erythrocyte preservation solution (0.005% glutaraldehyde and 0.05% paraformaldehyde)
It should be understood that the above examples are only for clarity of illustration and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications therefrom are within the scope of the invention.
Claims (10)
1. An erythrocyte preservation reagent, which is characterized in that the components of the erythrocyte preservation reagent comprise a solvent, glutaraldehyde and paraformaldehyde; the mass fraction of the glutaraldehyde in the solvent is 0.004-0.006%; the mass fraction of the paraformaldehyde in the solvent is 0.04-0.06%.
2. An erythrocyte preservation reagent according to claim 1, wherein the glutaraldehyde is present in the solvent at a mass fraction of 0.005%; the mass fraction of the paraformaldehyde in the solvent is 0.05%.
3. An erythrocyte preservation reagent according to claim 1 or 2, wherein the solvent is a solution containing trisodium citrate, potassium dihydrogen phosphate, disodium hydrogen phosphate dodecahydrate, sodium chloride, glucose, sucrose, adenine, creatinine, kanamycin, chloramphenicol, glutaraldehyde and/or paraformaldehyde.
4. A method of preserving red blood cells, said method comprising: erythrocytes are preserved by adding erythrocytes to the erythrocyte preservation reagent according to any one of claims 1 to 3.
5. A method of preserving red blood cells according to claim 4, wherein said method comprises: erythrocytes are preserved by adding erythrocytes to the erythrocyte preservation reagent according to any one of claims 1 to 3 to a mass fraction of 1 to 10%.
6. A method of preserving red blood cells according to claim 4 or 5, wherein said method comprises: red blood cells are added to the red blood cell preservation reagent according to claim 2 or 3 to a mass fraction of 5% to preserve the red blood cells.
7. Use of an erythrocyte preservation reagent according to any one of claims 1 to 3 or a method of preserving erythrocytes according to any one of claims 4 to 6 for preserving erythrocytes.
8. A red blood cell preservation solution obtained by preserving red blood cells by the method for preserving red blood cells according to any one of claims 4 to 6; the components of the erythrocyte preservation solution comprise a solvent, glutaraldehyde, paraformaldehyde and erythrocytes; the mass fraction of the glutaraldehyde in the solvent is 0.004-0.006%; the mass fraction of the paraformaldehyde in the solvent is 0.04-0.06%; the mass fraction of the red blood cells in the solvent is 1-10%.
9. The erythrocyte preservation solution of claim 8, wherein the glutaraldehyde is present in the solvent at a mass fraction of 0.005%; the mass fraction of the paraformaldehyde in the solvent is 0.05 percent; the mass fraction of the red blood cells in the solvent is 5%.
10. Use of the red blood cell preservation reagent according to any one of claims 1 to 3 or the method for preserving red blood cells according to any one of claims 4 to 6 or the red blood cell preservation solution according to claim 8 or 9 for blood group retrotyping detection, blood group antibody titer determination or red blood cell antibody screening and identification, characterized in that said use is for non-disease diagnostic and therapeutic purposes.
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Citations (3)
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| CN101285841A (en) * | 2008-04-25 | 2008-10-15 | 四川省迈克科技有限责任公司 | Three classifications whole blood quality control substance simulant and method for making same |
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| CN106614525A (en) * | 2016-12-19 | 2017-05-10 | 深圳市麦瑞科林科技有限公司 | Cell preservation solution as well as preparation method and use method thereof |
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| CN101285841A (en) * | 2008-04-25 | 2008-10-15 | 四川省迈克科技有限责任公司 | Three classifications whole blood quality control substance simulant and method for making same |
| CN102466730A (en) * | 2010-11-19 | 2012-05-23 | 南京神州英诺华医疗科技有限公司 | Novel method for matching blood reverse typing reagent |
| CN106614525A (en) * | 2016-12-19 | 2017-05-10 | 深圳市麦瑞科林科技有限公司 | Cell preservation solution as well as preparation method and use method thereof |
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