CN112684190A - ABO blood type positive typing and RhD blood type detection card and preparation method thereof - Google Patents
ABO blood type positive typing and RhD blood type detection card and preparation method thereof Download PDFInfo
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Abstract
The application discloses an ABO blood type positive typing and RhD blood type detection card and a preparation method thereof, and relates to the field of blood type typing cards. The blood type detection card comprises eight microcolumn gel tubes, wherein the eight microcolumn gel tubes comprise two gel tubes containing a monoclonal antibody A with IgM property, two gel tubes containing a monoclonal antibody B with IgM property, two gel tubes containing a monoclonal antibody D with IgM property and two blank gel tubes; the gel comprises gel swelling liquid and polyacrylamide glucan, wherein the gel swelling liquid comprises the following components: sodium dihydrogen phosphate, disodium hydrogen phosphate, NaCl, heparin sodium, polyalcohol, cyclodextrin and mannosyl glyceric acid. The blood type detection card prepared by the application can accurately detect ABO/RhD blood types of a person to be detected, and has a longer use validity period.
Description
Technical Field
The application relates to the field of blood type typing cards, in particular to an ABO blood type positive typing and RhD blood type detection card and a preparation method thereof.
Background
The ABO blood group system and the Rh blood group system are the two most important blood group systems of human, the high immunogenicity enables the ABO blood group system and the Rh blood group system to have important clinical significance, the ABO blood group system and the Rh blood group system are indispensable in emergency treatment, surgical treatment, anemia treatment, examination before blood transfusion and other processes, and the correctness of a test result is directly related to the life safety and danger of a patient.
Blood types are classified into A type, B type, AB type and O type according to the existence of A antigen and/or B antigen on the surface of erythrocyte membrane. The importance of the Rh blood group system in transfusion medicine is second to that of the ABO blood group system, the main antigens of the Rh system are D, C, C, E and E antigens, and because the immunogenicity of the D antigen is far stronger than that of other Rh antigens, the Rh blood groups of people are generally classified into two types of Rh negative and Rh positive according to the existence of the D antigen in clinic.
At present, a plurality of ABO blood type positive typing and RhD blood type detection card products are available on the market, but in most products, the sephadex is swelled by normal saline, after the swelled sephadex is mixed with an antibody, the stability of the antibody can not be maintained for a long time, and after the products are placed for a long time, the accuracy of detection results is obviously reduced.
Disclosure of Invention
The application provides an ABO blood group positive typing and RhD blood group detection card and a preparation method thereof, aiming at the problem that the stability of antibodies stored in the ABO blood group positive typing and RhD blood group detection card for a long time is not high in the prior art.
In a first aspect, the present application provides an ABO blood type is just stereotyped and RhD blood type detects card, can realize through following technical scheme:
an ABO blood group orthotyping and RhD blood group detection card comprises eight microcolumn gel tubes, wherein the eight microcolumn gel tubes comprise two gel tubes containing a monoclonal antibody A with IgM property, two gel tubes containing a monoclonal antibody B with IgM property, two gel tubes containing a monoclonal antibody D with IgM property and two blank gel tubes; the gel comprises gel swelling liquid and polyacrylamide glucan, wherein the gel swelling liquid comprises the following components:
0.1-0.4g/L of sodium dihydrogen phosphate;
0.1-0.4g/L of disodium hydrogen phosphate;
NaCl 1-2 g/L；
3-5 g/L of heparin sodium;
5-8g/L of polyhydric alcohol;
1-2 g/L of cyclodextrin;
mannosyl glyceric acid 0.05-0.08 g/L.
Through adopting above-mentioned technical scheme, the gel swelling liquid of this application includes buffer component, salt, anticoagulant and antibody stabilizer, and the combination of above components not only can be fine swelling dextran, makes the gel granule be the ball granule to better interception agglutination or sensitized erythrocyte improves the sensitivity that detects, can also improve the stability of antibody, makes the antibody keep for a long time under higher active state. Specifically, the buffer component can keep the whole system in a required buffer range, and provides the most suitable pH for the preservation of the antibody; the application further adds salt with lower concentration into the buffer system, which can fully swell the glucan, so that agglutinated red blood cells are easier to be trapped on the colloid, and unagglutinated red blood cells are easier to pass through the gaps between gel particles and reach the bottom of the gel tube, so that the result is easier to be read; the addition of the anticoagulant can improve the stability of the antibody on one hand, and can also improve the stability of red blood cells in a sample to be detected on the other hand, so that the accuracy of a detection result is improved; the three antibody stabilizers are compounded, so that the stability of physical properties, chemical properties and biological properties of the antibody can be effectively maintained, and great difference can not occur along with the prolonging of the storage time and the change of conditions, so that the accuracy of a detection result is ensured, and the service life of the detection card is prolonged.
Optionally, the gel swelling solution comprises the following components:
0.2-0.3g/L of sodium dihydrogen phosphate;
0.2-0.3g/L of disodium hydrogen phosphate;
NaCl 1.4-1.6 g/L；
heparin sodium 3.5-4.5 g/L;
6-7g/L of polyol;
1.5-1.8 g/L of cyclodextrin;
mannosyl glycerate 0.06-0.07 g/L.
Through adopting above-mentioned technical scheme, this application further prescribes a limit to the quantity of each component of gel swelling liquid to improvement testing result's that can be better accuracy, and the life of extension blood group test card.
Optionally, the polyhydric alcohol is sorbitol or mannitol.
By adopting the technical scheme, sorbitol or mannitol is selected as the polyhydric alcohol, the sorbitol or mannitol can better improve the stability of the antibody, and the two components are low in price, so that the cost of the product is reduced.
Optionally, the gel swelling solution further comprises 10-15mg/L of preservative.
Through adopting above-mentioned technical scheme, antiseptic is further added in gel swelling liquid in this application, and antiseptic addition can prevent breeding of antibody in long-term storage in-process bacterium, avoids forming the protein that combines with the antibody, has reduced false positive incidence to the accuracy of testing result has been improved.
Optionally, the preservative is thimerosal or glyceryl monocaprylate.
By adopting the technical scheme, the thimerosal or the glyceryl monocaprylate have strong resistance to microorganisms, have wide antimicrobial spectrum and can effectively inhibit the breeding of harmful microorganisms in a gel tube.
In a second aspect, the present application provides a method for preparing an ABO blood type positive typing and RhD blood type detection card, which is implemented by the following technical scheme:
a preparation method of an ABO blood group positive typing and RhD blood group detection card comprises the following steps:
s1, preparing a gel swelling solution: weighing specified amounts of sodium dihydrogen phosphate, disodium hydrogen phosphate, NaCl, heparin sodium, polyalcohol, cyclodextrin and mannosyl glyceric acid, and dissolving in water to obtain gel swelling solution;
s2, preparing gel: soaking the polyacrylamide glucan powder in 4-6 times of the volume of the gel swelling solution prepared in the step S1 for 24-48h, then washing for 3-5 times, removing gel breakage fragments and aggregated gel particles, and collecting the gel particles which are uniform in particle size and complete in spherical shape to obtain glucan gel;
s3, antibody dilution: respectively dissolving the monoclonal antibody A with the IgM property, the monoclonal antibody B with the IgM property and the monoclonal antibody D with the IgM property by using normal saline, so that the titer of the monoclonal antibody A with the IgM property is more than or equal to 128, the titer of the monoclonal antibody B with the IgM property is more than or equal to 128 and the titer of the monoclonal antibody D with the IgM property is more than or equal to 64;
s4, preparation of gel: mixing the three antibodies diluted in the step S3 with the sephadex prepared in the step S2 according to the volume ratio of (1-5) to 1, and respectively preparing gel containing IgM property monoclonal antibody A, IgM property monoclonal antibody B and IgM property monoclonal antibody D;
s5, subpackaging: and (4) respectively adding the three gels with the antibodies prepared in the step S4 and the gel prepared in the step S2 into different microcolumn tubes, and subpackaging two tubes for each gel to form the ABO blood type positive typing and RhD blood type detection cards of the eight microcolumn gel tubes.
By adopting the technical scheme, the gel swelling solution prepared by the method is used for swelling polyacrylamide glucan, then the diluted antibody and the swollen polyacrylamide glucan are mixed to obtain the gel containing the antibody, and the gel containing the antibody is added into a gel tube for sealing. The preparation method of the blood type detection card is simple, the blood type detection card is suitable for large-scale production, the prepared blood type detection card is high in specificity, the detection result is accurate, and the requirement of clinical rapid detection is met. In addition, the blood type detection card also has longer use time effect, and the market competitiveness of the product is improved.
Optionally, in step S1, the pH of the gel swelling solution is adjusted to 6.6 to 6.8.
By adopting the technical scheme, the pH value of the gel swelling solution is adjusted to 6.6-6.8, so that the polyacrylamide glucan is better swelled, and the antibody has better stability under the pH condition, so that the activity of the antibody is improved, and the accuracy of a detection result is improved.
Optionally, in step S2, the particle size of the polyacrylamide dextran is 20-50 nm.
By adopting the technical scheme, the particle size of the polyacrylamide glucan is limited to 20-50nm, and agglutinated red blood cells can be better intercepted, so that the detection result is easier to interpret.
Optionally, in step S2, the soaked polyacrylamide dextran is washed with the gel swelling solution for 1-2 times, and then washed with distilled water for 2-3 times.
Through adopting above-mentioned technical scheme, this application is washed the polyacrylamide dextran of swelling at first adopts gel swelling liquid, adopts distilled water to wash again, can effectively get rid of the gel particle of the damaged piece of gel and gathering to better entrapment agglutinate erythrocyte.
Optionally, in step S5, each micro-column tube is filled with 20-25 μ L of gel.
In summary, the present application has the following beneficial effects:
1. the method and the device can accurately detect the ABO/RhD blood type of the person to be detected, reduce the incidence rate of danger in the cross matching process, have simple detection process, can obtain the detection result in a short time, and improve the clinical detection efficiency;
2. the gel swelling solution can fully swell glucan on one hand, improves the detection sensitivity, can keep higher activity of an antibody for a longer time on the other hand, improves the accuracy and reliability of a detection result, and prolongs the service life of a product;
3. the detection card is simple in preparation method and suitable for large-scale production.
Detailed Description
The present application will be described in further detail with reference to examples.
Heparin sodium of the present application was purchased from Shanghai Allantin Biotechnology Ltd;
mannosyl glycerate of the present application was purchased from ibixin (shanghai) biotechnology limited;
the thimerosal of the present application is purchased from Shanxi Dadu pharmaceutical chemical Co., Ltd;
the present application relates to Qingdao Hua Yuan food additives Co., Ltd.
Preparation example 1
Preparing a gel swelling solution:
0.1g of sodium dihydrogen phosphate, 0.4g of disodium hydrogen phosphate, 1g of NaCl, 5g of heparin sodium, 5g of sorbitol, 2g of cyclodextrin and 0.05g of mannosylglyceric acid are weighed and dissolved in 1L of water, and the pH of the solution is adjusted to 6.8, so that the gel swelling solution is obtained.
Preparation example 2
Preparing a gel swelling solution:
0.4g of sodium dihydrogen phosphate, 0.1g of disodium hydrogen phosphate, 2g of NaCl, 3g of heparin sodium, 8g of mannitol, 1g of cyclodextrin and 0.08g of mannosylglyceric acid are weighed and dissolved in 1L of water, and the pH of the solution is adjusted to 6.6, so that the gel swelling solution is obtained.
Preparation example 3
Preparing a gel swelling solution:
0.2g of sodium dihydrogen phosphate, 0.3g of disodium hydrogen phosphate, 1.4g of NaCl, 4.5 g of heparin sodium, 6g of sorbitol, 1.8g of cyclodextrin and 0.06g of mannosylglyceric acid were weighed and dissolved in 1L of water, and the pH of the solution was adjusted to 6.7 to obtain a gel-swollen solution.
Preparation example 4
Preparing a gel swelling solution:
0.3g of sodium dihydrogen phosphate, 0.2g of disodium hydrogen phosphate, 1.6g of NaCl, 3.5 g of heparin sodium, 7g of mannitol, 1.5g of cyclodextrin and 0.07g of mannosylglyceric acid were weighed and dissolved in 1L of water, and the pH of the solution was adjusted to 6.7 to obtain a gel-swollen solution.
Preparation example 5
The preparation of the gel swelling solution is different from that of the preparation example 1 in that: thimerosal in an amount of 10mg was also added.
Preparation example 6
The preparation of the gel swelling solution is different from that of the preparation example 2 in that: also, 15mg of thimerosal was added.
Preparation example 7
The preparation of the gel swelling solution is different from that of the preparation example 3 in that: 10mg of glycerol monocaprylate were also added.
Preparation example 8
The preparation of the gel swelling solution is different from that of the preparation example 4 in that: 15mg of glycerol monocaprylate were also added.
Example 1
A preparation method of an ABO blood group positive typing and RhD blood group detection card comprises the following steps:
s1, preparing gel: soaking polyacrylamide glucan powder in 4 times of the volume of the gel swelling solution prepared in preparation example 1 for 48h, then washing with the gel swelling solution for 2 times, then washing with distilled water for 2 times, removing gel breakage fragments and aggregated gel particles, and collecting the gel particles which are uniform in particle size and completely spherical to obtain glucan gel; the particle size of the polyacrylamide glucan is 20-50 nm;
s2, antibody dilution: respectively dissolving the monoclonal antibody A with the IgM property, the monoclonal antibody B with the IgM property and the monoclonal antibody D with the IgM property by using normal saline, so that the titer of the monoclonal antibody A with the IgM property is more than or equal to 128, the titer of the monoclonal antibody B with the IgM property is more than or equal to 128 and the titer of the monoclonal antibody D with the IgM property is more than or equal to 64;
s3, preparing gel: mixing the three antibodies diluted in the step S2 with the sephadex prepared in the step S1 according to the volume ratio of 5:1, and respectively preparing gel containing the monoclonal antibody A with the IgM property, gel containing the monoclonal antibody B with the IgM property and gel containing the monoclonal antibody D with the IgM property;
s4, subpackaging: adding the three gels with the antibodies prepared in the step S3 and the gels prepared in the step S1 into different microcolumn tubes respectively, subpackaging two tubes for each gel, and subpackaging 25 mu L of the gel for each microcolumn tube to form an ABO blood type orthotyping and RhD blood type detection card with eight microcolumn gel tubes;
s5, sealing: the ABO blood type positive typing and RhD blood type test card prepared in this example was heat sealed with an aluminum foil composite sealing film.
Example 2
A preparation method of an ABO blood group positive typing and RhD blood group detection card comprises the following steps:
s1, preparing gel: soaking polyacrylamide glucan powder in 6 times of the volume of the gel swelling solution prepared in preparation example 1 for 24 hours, then washing the soaked polyacrylamide glucan powder for 1 time by using the gel swelling solution, then washing the soaked polyacrylamide glucan powder for 3 times by using distilled water, removing gel breakage fragments and aggregated gel particles, and collecting the gel particles which are uniform in particle size and complete in spherical shape to obtain glucan gel; the particle size of the polyacrylamide glucan is 20-50 nm;
s2, antibody dilution: respectively dissolving the monoclonal antibody A with the IgM property, the monoclonal antibody B with the IgM property and the monoclonal antibody D with the IgM property by using normal saline, so that the titer of the monoclonal antibody A with the IgM property is more than or equal to 128, the titer of the monoclonal antibody B with the IgM property is more than or equal to 128 and the titer of the monoclonal antibody D with the IgM property is more than or equal to 64;
s3, preparing gel: mixing the three antibodies diluted in the step S2 with the sephadex prepared in the step S1 according to the volume ratio of 1:1, and respectively preparing gel containing the monoclonal antibody A with the IgM property, gel containing the monoclonal antibody B with the IgM property and gel containing the monoclonal antibody D with the IgM property;
s4, subpackaging: adding the three gels with the antibodies prepared in the step S3 and the gels prepared in the step S1 into different microcolumn tubes respectively, and subpackaging two tubes for each gel and 20 mu L of gel for each microcolumn tube to form an ABO blood type orthotyping and RhD blood type detection card of eight microcolumn gel tubes;
s5, sealing: the ABO blood type positive typing and RhD blood type test card prepared in this example was heat sealed with an aluminum foil composite sealing film.
Example 3
A preparation method of an ABO blood group positive typing and RhD blood group detection card is different from that of the embodiment 1 in that: in step S1, polyacrylamide glucan powder was soaked with the gel-swelling solution prepared in preparation example 2.
Example 4
A method for preparing ABO blood group positive typing and RhD blood group detection card, which is different from the embodiment 2 in that: in step S1, polyacrylamide glucan powder was soaked with the gel-swelling solution prepared in preparation example 2.
Example 5
A preparation method of an ABO blood group positive typing and RhD blood group detection card is different from that of the embodiment 1 in that: in step S1, polyacrylamide glucan powder was soaked in the gel-swelling solution prepared in preparation example 3.
Example 6
A method for preparing ABO blood group positive typing and RhD blood group detection card, which is different from the embodiment 2 in that: in step S1, polyacrylamide glucan powder was soaked in the gel-swelling solution prepared in preparation example 3.
Example 7
A preparation method of an ABO blood group positive typing and RhD blood group detection card is different from that of the embodiment 1 in that: in step S1, polyacrylamide glucan powder was soaked in the gel-swelling solution prepared in preparation example 4.
Example 8
A method for preparing ABO blood group positive typing and RhD blood group detection card, which is different from the embodiment 2 in that: in step S1, polyacrylamide glucan powder was soaked in the gel-swelling solution prepared in preparation example 4.
Example 9
A preparation method of an ABO blood group positive typing and RhD blood group detection card is different from that of the embodiment 1 in that: in step S1, polyacrylamide glucan powder was soaked with the gel-swelling solution prepared in preparation example 5.
Example 10
A method for preparing ABO blood group positive typing and RhD blood group detection card, which is different from the embodiment 2 in that: in step S1, polyacrylamide glucan powder was soaked with the gel-swelling solution prepared in preparation example 5.
Example 11
A preparation method of an ABO blood group positive typing and RhD blood group detection card is different from that of the embodiment 1 in that: in step S1, polyacrylamide glucan powder was soaked in the gel-swelling solution prepared in preparation example 6.
Example 12
A method for preparing ABO blood group positive typing and RhD blood group detection card, which is different from the embodiment 2 in that: in step S1, polyacrylamide glucan powder was soaked in the gel-swelling solution prepared in preparation example 6.
Example 13
A preparation method of an ABO blood group positive typing and RhD blood group detection card is different from that of the embodiment 1 in that: in step S1, polyacrylamide glucan powder was soaked with the gel-swelling solution prepared in preparation example 7.
Example 14
A method for preparing ABO blood group positive typing and RhD blood group detection card, which is different from the embodiment 2 in that: in step S1, polyacrylamide glucan powder was soaked with the gel-swelling solution prepared in preparation example 7.
Example 15
A preparation method of an ABO blood group positive typing and RhD blood group detection card is different from that of the embodiment 1 in that: in step S1, polyacrylamide glucan powder was soaked with the gel-swelling solution prepared in preparation example 8.
Example 16
A method for preparing ABO blood group positive typing and RhD blood group detection card, which is different from the embodiment 2 in that: in step S1, polyacrylamide glucan powder was soaked with the gel-swelling solution prepared in preparation example 8.
Comparative example 1
A preparation method of an ABO blood group positive typing and RhD blood group detection card is different from that of the embodiment 1 in that: the gel swelling solution does not contain heparin sodium.
Comparative example 2
A preparation method of an ABO blood group positive typing and RhD blood group detection card is different from that of the embodiment 1 in that: the gel swelling solution does not contain cyclodextrin.
Comparative example 3
A preparation method of an ABO blood group positive typing and RhD blood group detection card is different from that of the embodiment 1 in that: the gel swelling solution does not contain mannosyl glyceric acid.
Comparative example 4
A preparation method of an ABO blood group positive typing and RhD blood group detection card is different from that of the embodiment 1 in that: the gel swelling solution is physiological saline.
A preparation method of an ABO blood group positive typing and RhD blood group detection card is different from that of the embodiment 1 in that:
the application method of the ABO blood group orthotyping and RhD blood group detection card prepared in the embodiments 1-16 and the comparative examples 1-4 is as follows:
1. the ABO blood group orthotyping and RhD blood group detection card comprises eight microcolumn gel tubes, wherein the first and fifth gel tubes are gel tubes containing a monoclonal antibody A with IgM property, the second and sixth gel tubes are gel tubes containing a monoclonal antibody B with IgM property, the third and seventh gel tubes are gel tubes containing a monoclonal antibody D with IgM property, and the fourth and eighth gel tubes are blank gel tubes;
2. setting red blood cells of a subject to be detected to have a concentration of 0.5-0.8% by using normal saline, and adding the red blood cells into the first to fourth microtubes or the fifth to eighth microtubes respectively, wherein each microtube contains 50 mu L of red blood cells;
3. centrifuging with a centrifuge special for a microcolumn gel card at 900rpm for 2min and 1500rpm for 3min, taking out, and judging the result with naked eyes;
4. and (4) interpretation of results: the result is negative when the red blood cells sink to the bottom of the gel of the microcolumn, the result is positive when the red blood cells float on the surface of the gel or in the gel, the specific interpretation standard is shown in table 1, and the judgment result is shown in table 2.
Note: "+" is positive and "0" is negative; if the Ctr tube shows positive, hemolytic or double results, the experimental result is invalid, and the reason needs to be further determined.
Performance test
The blood type detection cards prepared in examples 1-16 and comparative examples 1-4 of the present application and the blood type detection cards produced in 18 months and 24 months were used to perform blood type detection on 100 blood volunteers of type A, 100 blood volunteers of type B, 100 blood volunteers of type AB, 100 blood volunteers of type O, 50 blood volunteers of type RhD positive, and 20 blood volunteers of type RhD negative, and the experimental results are shown in tables 3-5.
As can be seen from tables 3-5, the blood type test card prepared in the embodiments 1-16 of the present application can not only test the ABO blood type, but also test the RhD blood type, the test result is the same as the predetermined result, and the test result is accurate and reliable. In addition, when the blood type detection card after 18 months of production is used for detecting a sample for determining the blood type, the blood type detection cards prepared in the embodiments 1 to 16 can still accurately detect the sample to be detected, especially the blood type detection cards prepared in the embodiments 9 to 16 detect the blood type of a person to be detected after 24 months of production, the accuracy of the experimental result is 100%, and the product has a longer effective period and higher accuracy.
The difference between the comparative examples 1-3 and the example 1 is that the gel swelling solution does not contain heparin sodium, cyclodextrin and mannosylglyceric acid, and as can be seen from tables 3-5, the blood type detection card prepared by the comparative examples 1-3 is placed for 18 months, and then the blood type of a person to be detected is detected, so that the accuracy of the detection result is obviously reduced, and the experimental results show that the heparin sodium, the cyclodextrin and the mannosylglyceric acid can maintain the long-term stability of the antibody, and the accuracy of the detection result is improved.
The difference between the comparative example 4 and the example 1 is that the physiological saline is used as the gel swelling agent, and as can be seen from tables 3 to 5, the blood type detection card prepared by the comparative example 4 is used for detecting the blood type of a person to be detected after being placed for 18 months, the accuracy of the detection result is very poor, and the above experiment results show that the physiological saline can not keep the stability of the antibody for a long time.
The embodiments of the present invention are preferred embodiments of the present application, and the scope of protection of the present application is not limited by the embodiments, so: all equivalent changes made according to the structure, shape and principle of the present application shall be covered by the protection scope of the present application.
Claims (10)
1. The utility model provides a card is examined to ABO blood group is just stereotyped and rhD blood group which characterized in that: the kit comprises eight microcolumn gel tubes, wherein the eight microcolumn gel tubes comprise two gel tubes containing a monoclonal antibody A with IgM property, two gel tubes containing a monoclonal antibody B with IgM property, two gel tubes containing a monoclonal antibody D with IgM property and two blank gel tubes; the gel comprises gel swelling liquid and polyacrylamide glucan, wherein the gel swelling liquid comprises the following components:
0.1-0.4g/L of sodium dihydrogen phosphate;
0.1-0.4g/L of disodium hydrogen phosphate;
NaCl 1-2 g/L；
3-5 g/L of heparin sodium;
5-8g/L of polyhydric alcohol;
1-2 g/L of cyclodextrin;
mannosyl glyceric acid 0.05-0.08 g/L.
2. The ABO blood group orthotyping and RhD blood group testing card of claim 1, wherein: the gel swelling solution comprises the following components:
0.2-0.3g/L of sodium dihydrogen phosphate;
0.2-0.3g/L of disodium hydrogen phosphate;
NaCl 1.4-1.6 g/L；
heparin sodium 3.5-4.5 g/L;
6-7g/L of polyol;
1.5-1.8 g/L of cyclodextrin;
mannosyl glycerate 0.06-0.07 g/L.
3. The ABO blood group orthotyping and RhD blood group test card of claim 1 or 2, wherein: the polyalcohol is sorbitol or mannitol.
4. The ABO blood group orthotyping and RhD blood group test card of claim 1 or 2, wherein: the gel swelling solution also comprises 10-15mg/L preservative.
5. The ABO blood group orthotyping and RhD blood group testing card according to claim 4, wherein: the antiseptic is selected from thimerosal or glyceryl monocaprylate.
6. A method of preparing the ABO blood group positive typing and RhD blood group testing card of any one of claims 1 to 5, comprising the steps of:
s1, preparing a gel swelling solution: weighing specified amounts of sodium dihydrogen phosphate, disodium hydrogen phosphate, NaCl, heparin sodium, polyalcohol, cyclodextrin and mannosyl glyceric acid, and dissolving in water to obtain gel swelling solution;
s2, preparing gel: soaking the polyacrylamide glucan powder in 4-6 times of the volume of the gel swelling solution prepared in the step S1 for 24-48h, then washing for 3-5 times, removing gel breakage fragments and aggregated gel particles, and collecting the gel particles which are uniform in particle size and complete in spherical shape to obtain glucan gel;
s3, antibody dilution: respectively dissolving the monoclonal antibody A with the IgM property, the monoclonal antibody B with the IgM property and the monoclonal antibody D with the IgM property by using normal saline, so that the titer of the monoclonal antibody A with the IgM property is more than or equal to 128, the titer of the monoclonal antibody B with the IgM property is more than or equal to 128 and the titer of the monoclonal antibody D with the IgM property is more than or equal to 64;
s4, preparation of gel: mixing the three antibodies diluted in the step S3 with the sephadex prepared in the step S2 according to the volume ratio of (1-5) to 1, and respectively preparing gel containing IgM property monoclonal antibody A, IgM property monoclonal antibody B and IgM property monoclonal antibody D;
s5, subpackaging: and (4) respectively adding the three gels with the antibodies prepared in the step S4 and the gel prepared in the step S2 into different microcolumn tubes, and subpackaging two tubes for each gel to form the ABO blood type positive typing and RhD blood type detection cards of the eight microcolumn gel tubes.
7. The method of claim 6, wherein in step S1, the pH of the gel swelling solution is adjusted to 6.6-6.8.
8. The method of claim 6, wherein in step S2, the particle size of the polyacrylamide dextran is 20-50 nm.
9. The method of claim 6, wherein in step S2, the soaked polyacrylamide dextran is washed with gel swelling solution for 1-2 times and then with distilled water for 2-3 times.
10. The method of claim 6, wherein each micro-column tube is filled with 20-25 μ L of gel in step S5.
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