CN221550705U - Rapid detection card for antibody screening and cross matching - Google Patents
Rapid detection card for antibody screening and cross matching Download PDFInfo
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- CN221550705U CN221550705U CN202322913548.2U CN202322913548U CN221550705U CN 221550705 U CN221550705 U CN 221550705U CN 202322913548 U CN202322913548 U CN 202322913548U CN 221550705 U CN221550705 U CN 221550705U
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- Investigating Or Analysing Biological Materials (AREA)
Abstract
The utility model relates to a rapid detection card for antibody screening and cross matching, which comprises a fixed plate, a supporting plate arranged on the top of the fixed plate and a reaction cavity arranged on the fixed plate, wherein the reaction cavity is opened on the supporting plate, a sealing film for sealing the opening of the reaction cavity is covered on the surface of the supporting plate, a glass bead layer is filled at the bottom end inside the reaction cavity, and an irregular rapid reaction coagulant for antibodies is arranged above the glass bead layer. According to the utility model, the antigen-antibody specific reaction principle, the irregular antibody rapid reaction coagulant technology and the filtration molecular sieve technology are organically combined together, and after red blood cells and/or serum which participate in the reaction are added, the reaction is incubated for 1min at 37 ℃, and the agglutination result can be observed by centrifugation, so that the method has the advantages of high sensitivity, simplicity, rapidness, stable and reliable result and great shortening of the detection time.
Description
Technical Field
The utility model relates to a rapid detection card for antibody screening and cross matching.
Background
Since Lapierre and the like in French in 1986 invented a micro-column gel card, the advantages of simplicity, rapidness, good repeatability, high sensitivity, easy standardization and automation and the like lead the micro-column gel card to be widely applied in the blood group serology field. The american society of transfusion technology has included this technology in the pre-transfusion compatibility test in the handbook of transfusion technology, 12 th edition (1996). Because the low-ion anti-globulin and anti-C3 d reagent is added in the kit as a reaction medium, the sephadex is used as a separation medium, the reaction process is carried out by placing the kit in an incubator at 37 ℃ for 15min, and then centrifuging the kit for 5min by using a special centrifuge to observe results, and the required time is longer, the kit is not applicable to emergency treatment of the hemorrhagic shock patient, and on the basis, the kit has important practical significance in researching a faster and safer domestic cross blood matching detection card.
Disclosure of utility model
The utility model aims to solve the technical problem of providing a rapid detection card for antibody screening and cross matching, which greatly shortens the detection time and improves the timeliness of transfusion treatment.
The technical scheme adopted by the utility model is as follows:
A quick detection card for antibody screening and cross matching is characterized in that the quick detection card comprises a fixed plate, a supporting plate arranged at the top of the fixed plate and a reaction cavity arranged on the fixed plate, wherein the reaction cavity is opened in the supporting plate, a sealing film used for sealing the opening of the reaction cavity is covered on the surface of the supporting plate, a glass bead layer is filled at the bottom end inside the reaction cavity, and an irregular quick reaction coagulant for antibodies is arranged above the glass bead layer.
Further, the cross section of the reaction cavity is V-shaped or U-shaped.
Further, the number of the reaction chambers is 6.
Further, the glass bead layer comprises a plurality of glass beads, and the particle size of the glass beads is 80-100 mu m.
Further, the volume ratio of the irregular antibody quick reaction coagulant to the glass bead layer is 1:1-2:1.
The utility model has the positive effects that:
According to the utility model, an antigen-antibody specific reaction principle, an irregular antibody rapid reaction coagulant technology and a filtration molecular sieve technology are organically combined together, glass beads are adopted as a separation medium, erythrocytes and/or serum which participate in the reaction are added, and then the reaction is incubated at 37 ℃ for 1min, and an agglutination result can be observed after centrifugation, so that the method has the advantages of high sensitivity, simplicity, rapidness, stable and reliable result, greatly shortened detection time, and is used for screening of blood samples of patients before blood transfusion and cross matching test, and the timeliness of blood transfusion treatment is improved. The reaction cavity adopts a V-shaped or U-shaped structure, and compared with the reaction cavity adopted in the traditional microcolumn gel card, the reaction cavity has simpler structure and lower processing cost.
Drawings
FIG. 1 is a schematic diagram of the structure of the present utility model;
FIG. 2 is a schematic view of the top opening of the reaction chamber of the present utility model;
FIG. 3 is a schematic diagram of another embodiment of the present utility model;
FIG. 4 shows the results of detection using a microcolumn gel card;
FIG. 5 shows the test results of a rapid test card employing the present utility model.
Detailed Description
As shown in figures 1 and 2, the utility model provides a rapid detection card for antibody screening and cross matching, which comprises a vertical fixing plate 1, a supporting plate 2 vertically arranged at the top of the fixing plate 1, six reaction chambers 4 arranged on the fixing plate 1 and a sealing film 3 attached to the supporting plate 2, wherein the cross section of the reaction chambers 4 is V-shaped, the top opening of the reaction chambers is arranged on the supporting plate 2, the sealing film 3 is attached to the flange of the opening of the reaction chambers 4 to seal the flange, the glass bead layers 5 consisting of a plurality of glass beads are filled at the bottom end in the reaction chambers 4, and an irregular rapid antibody reaction coagulant 6 is arranged above the glass bead layers 5.
Preferably, the glass beads have a particle size of 80 to 100 μm. And the volume ratio of the irregular antibody quick reaction coagulant 6 to the glass microsphere layer 5 is 1:1-2:1.
As shown in fig. 1 and 2, in this embodiment, the cross section of the reaction chamber 4 is V-shaped. In another embodiment, the reaction chamber 4 may also have a U-shaped cross section, as shown in FIG. 3.
The test principle of the utility model is as follows:
The antigen-antibody specific reaction principle, the irregular antibody rapid reaction technology and the filtration molecular sieve technology are organically combined together. Under normal physiological conditions, sialic acid on the surface of erythrocytes has a large amount of negative charge, a Zeta potential is formed, and the repulsive interaction generated by the Zeta potential keeps a certain distance between erythrocytes, so that erythrocytes can flow in blood vessels without aggregation, but the distance is far greater than the distance between two Fab ends of an IgG antibody, and therefore, even if the surface of erythrocytes is sensitized by irregular antibodies, agglutination reaction can not occur. The low ion solution (with the ionic strength of 0.01 mol/L) in the irregular antibody quick reaction coagulant 6 can reduce the ionic strength of plasma and erythrocyte reaction medium from 0.17mol/L to 0.03mol/L, can promote the combination of erythrocyte and antibody in the plasma, quicken the reaction speed, lead the IgG antibody to finish erythrocyte sensitization in 1min under the room temperature condition, lead protamine and polybrene to be macromolecule groups with positive charges, generate a large number of cations after dissolution, can neutralize the negative charge carried by sialic acid on the surface of the erythrocyte, lead the Zeta potential of the surface of the erythrocyte to be reduced, shorten the distance between cells, be favorable for the specific agglutination reaction of the IgG antibody of small molecules and the corresponding antigen on the surface of the erythrocyte through the bridging effect, lead sodium oxalate to neutralize the excessive positive charge of the protamine through the ionic effect, eliminate the nonspecific reaction, lead the PEG to be a polymer of neutral glycol without charges, improve the relative concentration of erythrocyte through the space rejection effect, lead the low concentration PEG to enhance the agglutination force, promote the agglutination reaction, and promote the occurrence of the detection sensitivity. BSA can increase the dielectric constant of the medium and promote the agglutination reaction of irregular antibodies. Glass beads with proper pore sizes are selected, gaps among the beads only allow single red blood cells to pass through, when irregular antibodies react with red blood cells of a donor, the red blood cells aggregated under the action of centrifugal force cannot pass through gaps among the beads and are blocked at the upper part or in the middle of dispersion of the beads, the result is positive, the positive or cross matching result of irregular antibody screening test is indicated, if serum of a patient does not react with the red blood cells of the donor, the red blood cells are not aggregated, and the red blood cells deposit at the bottom of a reaction hole after centrifugation, and the result is negative. And (3) according to the comprehensive analysis of the detection results, judging whether meaningful reactive antibodies exist in the patient body or are matched with the cross matching of the donor.
The manufacturing steps of the utility model are as follows:
Step one, preparing blank detection cards, wherein the blank detection cards are made of hard plastic materials and have the specification of 70mm multiplied by 55mm, and each detection card is provided with 6 reaction cavities.
And step two, preparing the irregular antibody quick reaction coagulant. Irregular antibody fast reaction coagulants are more well known in the published literature, and in this embodiment, these existing irregular antibody fast reaction coagulants may be selected. The utility model also discloses an irregular antibody quick reaction coagulant selected in the embodiment, which comprises protamine sulfate (0.1%), PEG (molecular weight 6000,2.5%), sodium oxalate (0.5%) and BSA (1%), wherein the solution with low ionic strength (5% glucose+0.2% EDTA-2 NA) is dissolved in distilled water to a final pH value of 6.2-6.5, and finally a proper amount of sodium azide is added for preservation (0.02%), so that the solution can be preserved for 1 year at 4 ℃.
And thirdly, selecting glass beads with the particle size of 80-100 mu m.
And step four, mixing the irregular antibody quick reaction coagulant with the glass beads according to the volume ratio of 1:1-2:1 to prepare the coagulant bead suspension perfusate. Adding 50 mu L of gel suspension into each tube, sucking the gel suspension into the bottom of a gel column by using a micropipette or adopting a 2mL syringe, centrifuging for 1min at 1500g to cause the microbeads to uniformly precipitate at the bottom of a reaction cavity, avoiding generating bubbles or faults in the column, and suspending an irregular antibody quick reaction coagulant medium above the microbeads to prepare the microbead detection card containing the irregular antibody coagulant.
And fifthly, sealing the upper opening of the reaction cavity by using aluminum foil paper for the microbead detection card in the step four through a film pressing mode, labeling, and then preserving the detection card at 18-25 ℃ for more than 1 year, and more preferably preserving at 4 ℃.
The experiment performed with the rapid test card of the present utility model is as follows:
1. Data and method
2023, 5-2023, 7, 178 Cross-matched blood specimens for censoring, 325 specimen screening by blood station gratuitous donor antibodies, all specimen being EDTA anticoagulated whole blood. The blood specimen is subjected to antibody screening and cross matching test by adopting a low-ion anti-globulin microcolumn Gel card method (DG Gel Coombs card) and the rapid detection card. And further carrying out antibody specificity identification on the specimen with positive detection, and comparing the consistency of the detection results of the specimen with the antibody specificity identification.
2. Antibody screening assays
2.1 Method for on-the-fly gel card
Taking 1 detection card, marking 3 holes of each sample to be detected as I, II and III, respectively adding 50 mu L of serum or plasma of the person to be detected into each marked hole, adding 50 mu L of corresponding 0.8-1% irregular antibody screening cell suspension into each hole according to the mark, vertically and transversely standing the detection card on a table top, uniformly mixing plasma and red blood cells in the card at the edge of the light buckle card, then placing the card in an incubator for incubation for 15min at 37 ℃, taking out the card, placing the card in a special centrifuge for gradient centrifugation for 5min, and observing and recording the agglutination result.
2.2 Rapid test card antibody screening assays of the utility model
Taking 50 mu L of serum or plasma of a person to be detected, adding 50 mu L of corresponding 0.8-1% irregular antibody screening cell suspension into each hole according to the mark, uniformly mixing, placing in an incubator for incubation for 1min at 37 ℃, taking out, placing in a centrifuge for centrifugation for 2min at 1500 rpm, and observing and recording the agglutination result.
3. The rapid detection card cross blood matching test of the utility model
Taking 1 sample of each patient, marking 2 holes, namely a primary side and a secondary side, adding 50 mu L of serum or plasma of the patient into the primary side holes, adding 50 mu L of red blood cell suspension of 0.8-1% of blood donor into the secondary side holes, adding 50 mu L of serum or plasma of the blood donor into the secondary side holes, uniformly mixing, incubating and centrifuging according to different requirements of the two cards, and observing and recording the agglutination result.
4. Results
4.1 Cross-matching results
178 Cases of cross matching experiments: the microcolumn gel detection card method and the rapid detection card method of the utility model have 4 cases of main side cross matching incompatibility and 5 cases of secondary side incompatibility respectively; the matching rate of the two methods is 100%, and the difference of the detected incomplete antibodies has no statistical significance (P is more than 0.05).
4.2 Antibody screening results
The 325 specimens were tested for antibody screening by two methods, and 9 specimens were positive in the results, and the consistency of the results of antibody screening was good. The difference between the two methods of detection of incomplete antibodies was not statistically significant (P > 0.05).
4.3 The same sample, the consistency of the detection results of the two methods is good, and the detection results are shown in fig. 4 and 5. (wherein, the reaction chamber in FIG. 5 has the same structure as that of the conventional reaction chamber of the microcolumn gel card, since no improvement has been made in the reaction chamber at the stage of the test)
The above embodiments are only for illustrating the technical solution of the present utility model, and are not limiting; although the utility model has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some of the technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit and scope of the technical solutions of the embodiments of the present utility model.
Claims (5)
1. A quick detection card for antibody screening and cross matching is characterized in that the quick detection card comprises a fixed plate (1), a supporting plate (2) arranged at the top of the fixed plate and a reaction cavity (4) arranged on the fixed plate (1), wherein the reaction cavity (4) is opened on the supporting plate (2), a sealing film (3) used for sealing the opening of the reaction cavity (4) is covered on the surface of the supporting plate (2), a glass bead layer (5) is filled at the bottom end inside the reaction cavity (4), and an irregular quick reaction coagulant (6) for antibodies is arranged above the glass bead layer (5).
2. A rapid test card for antibody screening and cross-matching according to claim 1, characterized in that the reaction chamber (4) is V-shaped or U-shaped in cross-section.
3. A rapid test card for antibody screening and cross-matching according to claim 1 or 2, characterized in that the number of reaction chambers (4) is 6.
4. The rapid detection card for antibody screening and cross matching according to claim 1, wherein the glass bead layer (5) comprises a plurality of glass beads, and the particle size of the glass beads is 80-100 μm.
5. The rapid detection card for antibody screening and cross-matching according to claim 1, wherein the volume ratio of the irregular antibody rapid reaction coagulant (6) to the glass bead layer (5) is 1:1-2:1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202322913548.2U CN221550705U (en) | 2023-10-30 | 2023-10-30 | Rapid detection card for antibody screening and cross matching |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202322913548.2U CN221550705U (en) | 2023-10-30 | 2023-10-30 | Rapid detection card for antibody screening and cross matching |
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| Publication Number | Publication Date |
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| CN221550705U true CN221550705U (en) | 2024-08-16 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN202322913548.2U Active CN221550705U (en) | 2023-10-30 | 2023-10-30 | Rapid detection card for antibody screening and cross matching |
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| Country | Link |
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| CN (1) | CN221550705U (en) |
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- 2023-10-30 CN CN202322913548.2U patent/CN221550705U/en active Active
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