CN211905397U - Human ABO blood type reverse-typing rapid detection test strip - Google Patents
Human ABO blood type reverse-typing rapid detection test strip Download PDFInfo
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- CN211905397U CN211905397U CN202020368044.0U CN202020368044U CN211905397U CN 211905397 U CN211905397 U CN 211905397U CN 202020368044 U CN202020368044 U CN 202020368044U CN 211905397 U CN211905397 U CN 211905397U
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Abstract
The utility model discloses a reverse quick test examination strip of stereotyping of people ABO blood group, including a examination strip body, including a base plate and locate a application of sample pad, an antigen pad, a reaction membrane and a water absorption pad on this base plate, the middle part of this base plate is located to the reaction membrane, the other end of the one end contact antigen pad of application of sample pad, the other end of the one end contact reaction membrane of antigen pad, the other end of the one end contact water absorption pad of reaction membrane. The utility model discloses make the anti-design of blood group detect more convenient, quick, and sensitivity is higher, and the specificity is better, effectively solves the anti-design of clinical unit blood group and detects complicacy, difficult scheduling problem.
Description
Technical Field
The utility model particularly relates to a reverse typing short-term test examination strip of people ABO blood type.
Background
The ABO blood group system classifies blood types into O, A, B and AB blood types according to the difference between the agglutinants A and B on erythrocytes. The A-type red blood cells contain an A antigen, and the blood serum contains an anti-B antibody; b antigen is contained on B type blood red cells, and anti-A antibody is contained in serum; the O-type red blood cells have no A and B antigens, and the serum contains anti-A and anti-B antibodies; AB type red blood cells contain two antigens, A and B, and serum does not contain anti-A and anti-B antibodies. The antigen A and the anti-A antibody, and the antigen B and the anti-B antibody can generate immune combination reaction, so that the erythrocyte is condensed. ABO blood group is determined by both erythrocyte surface antigens and antibodies in serum. Detecting erythrocyte antigens by using standard anti-A and anti-B antibodies, which is called positive typing; antibodies in serum were detected using standard A and B antigens, called retrotyping. The ABO blood type can be correctly judged only by detecting the positive and negative typing simultaneously.
Correct blood grouping is an important task for ensuring clinical transfusion safety and life saving, ABO blood grouping is one of the routine pre-transfusion serological testing items, and in order to ensure correct blood grouping, the ministry of health was specifically regulated in 2000 letters (health and medical act [2000] 184): each blood-supplying and receiving individual should be subjected to positive and negative ABO blood type typing. The most commonly used ABO blood type anti-typing identification method in clinical laboratories at present is a hemagglutination test, the used standard red blood cells of type A, type B and type O have short storage life, are generally stored for 3 months at 2-8 ℃, and are easy to cause microbial contamination and hemolysis, if the blood cells are prepared by blood units, the preparation methods of all units are different due to the lack of unified standards, the operation is complicated, the quality of an anti-typing red blood cell reagent is extremely unstable due to the difference of the components and physicochemical properties of red blood cell preservation liquid, the storage life in vitro is short, a centrifugal device or a glass slide is also needed during reaction, the operation is complicated, the result interpretation is difficult, the identification error is easy to cause, and even serious transfusion accidents occur.
CN103033632A discloses an ABO blood group reverse typing colloidal gold kit and a preparation method thereof, which solves the problem of short storage life of fresh red blood cells by preparing red blood cell membrane blood group antigen extract and then carrying out colloidal gold labeling. The principle is that erythrocyte surface blood group antigen is coated on the surface of the particle, so that the particle has ABO blood group antigenicity and can generate specific immunoreaction with corresponding antibody in serum, and after slight oscillation, if agglutination occurs, specific antibody-antigen complex is generated, and the reaction is positive reaction; if agglutination does not occur, it indicates that no antigen-antibody complex is formed, and the reaction is negative. Because the blood group antigens used by the kit are only fragments of primarily broken erythrocyte membranes and are not purified, the data show that the blood group antigens of the erythrocyte membranes are more than 200, so that the reverse typing of ABO blood group identification by specifically detecting anti-A and anti-B cannot be ensured, and the vibration reaction is carried out by auxiliary equipment.
The technical scheme disclosed in CN107290520A is also immunochromatography detection reverse typing, wherein labeled anti-human IgM antibody and anti-human IgG antibody are fixed on glass fiber, a conjugate of a and B antigens is respectively coated on a cellulose membrane to prepare two detection lines, and then the two detection lines are assembled into a test strip to detect blood type antibodies in a sample, such as anti-a and anti-B, namely: the kit comprises marked anti-human IgM antibody, anti-human IgG antibody, antibodies to be detected (anti-A and anti-B) and coating antigen (A and B antigen conjugate), and is characterized in that the anti-human IgM antibody and the anti-human IgG antibody can be combined with all IgM and IgG type antibodies in a sample, and the antibodies in human blood are very complex and various, wherein the blood type anti-A and anti-B are only a small part of the blood type anti-A and anti-B, so that non-specific combination is easily caused, and the sensitivity is low. Meanwhile, the blood group antigen is an artificially synthesized antigen, the actual application of the artificially synthesized antigen is not reported at present, and the artificially synthesized antigen has the possibility of insufficient affinity with the blood group antibody in a human body, so that the detection sensitivity is easy to fail to meet the requirement.
SUMMERY OF THE UTILITY MODEL
An object of the utility model is to overcome prior art defect, provide a reverse design quick test examination strip of people ABO blood type.
The technical scheme of the utility model as follows:
a rapid test strip for reverse typing of human ABO blood type comprises:
the test strip body comprises a substrate, and a sample adding pad, an antigen pad, a reaction film and a water absorbing pad which are arranged on the substrate, wherein the reaction film is arranged in the middle of the substrate, one end of the sample adding pad is contacted with the other end of the antigen pad, one end of the antigen pad is contacted with the other end of the reaction film, and one end of the reaction film is contacted with the other end of the water absorbing pad;
purified erythrocyte blood group A antigen and erythrocyte blood group B antigen marked by tracer particles are fixed on the antigen pad through physical adsorption,
the reaction membrane is provided with a detection area, the detection area is sequentially provided with a first detection line, a second detection line and a contrast line from the other end to one end of the reaction membrane, the first detection line is coated with a solidified anti-A antibody, the second detection line is coated with a solidified anti-B antibody, and the contrast line is coated with a solidified anti-RBC antibody.
In a preferred embodiment of the present invention, the other end of the antigen pad is overlapped on one end of the sample addition pad.
In a preferred embodiment of the present invention, one end of the antigen pad is overlapped on the other end of the reaction membrane.
In a preferred embodiment of the present invention, the other end of the absorbent pad is overlapped on one end of the reaction membrane.
In a preferred embodiment of the present invention, the tracer particles are colloidal gold.
The utility model discloses an in a preferred embodiment, still include one and go up the casing, should go up the casing and locate on the base plate and cover application of sample pad, antigen pad, reaction membrane and absorbent pad should go up the casing and have the observation window that the application of sample hole and a detection zone that corresponds the reaction membrane of a corresponding application of sample pad.
The utility model has the advantages that:
1. the utility model discloses make the anti-design of blood group detect more convenient, quick, and sensitivity is higher, and the specificity is better, effectively solves the anti-design of clinical unit blood group and detects complicacy, difficult scheduling problem.
2. The purified erythrocyte blood group A antigen and the erythrocyte blood group B antigen marked by the tracer particles are fixed on the antigen pad of the utility model through physical adsorption, can be specifically and immunologically combined with the corresponding anti-A/B blood group antibody, and the specificity is better; because the antigen is naturally present in the human body, the affinity with the blood-type antibody in the human body is better, and the sensitivity of antigen-antibody binding is higher.
3. The utility model discloses a detection zone is equipped with a first detection line, a second detection line and a control line in proper order from the other end of reaction film to one end, and the anti-anti A antibody of coating solidification on the first detection line, the anti-anti B antibody of coating solidification on the second detection line, and the anti RBC antibody of coating solidification on the control line can guarantee that the immunity of specificity and corresponding human blood type antibody takes place to combine, effectively shields the interference of other non-anti A, anti B antibody in the sample, and sensitivity is higher.
Drawings
Fig. 1 is a schematic structural diagram of embodiment 1 of the present invention.
Fig. 2 is a schematic structural diagram of an upper housing according to embodiment 1 of the present invention.
Detailed Description
The technical solution of the present invention will be further illustrated and described by the following specific embodiments.
Example 1
A rapid test strip for reverse typing of human ABO blood type comprises a test strip body 1 and an upper shell 2.
As shown in fig. 1, the test strip body 1 includes a substrate 10, and a sample addition pad 11, an antigen pad 12, a reaction membrane 13 and a water absorption pad 14 disposed on the substrate 10, wherein the reaction membrane 13 is disposed in the middle of the substrate 10, the other end of the antigen pad 12 is connected to one end of the sample addition pad 11, one end of the antigen pad 12 is connected to the other end of the reaction membrane 13, and the other end of the water absorption pad 14 is connected to one end of the reaction membrane 13;
the antigen pad 12 is fixed with erythrocyte blood group A antigen and erythrocyte blood group B antigen which are marked and purified by colloidal gold through physical adsorption, and the preparation method of the A antigen and the B antigen comprises the following steps:
membrane antigen is extracted by human erythrocyte crushing method, hypotonic swelling crushing is adopted, and the component of hypotonic lysis solution is 0.1% (g/L) CaCl20.1% (g/L) Triton X-100, 0.1% (g/L) SDS and 0.1% (g/L) EDTA2K at PH 7.4, comprising the steps of:
(1) washing: washing fresh human red blood cells with physiological saline, centrifuging at 2000rpm for 3min, removing supernatant, repeating for three times, and collecting red blood cells;
(2) crushing: adding lysis solution into washed erythrocytes and lysis solution according to the volume ratio of 4: 1, mixing uniformly, immediately adding 1mM dimethyl sulfoxide, wherein the volume ratio of dimethyl sulfoxide to erythrocytes is 95: 4, mixing uniformly, and standing for 30 min;
(3) collecting: centrifuging at 15000rpm for 10min, collecting supernatant, wherein A, B blood group antigen has been dissolved in the supernatant;
(4) absorption: adding an anti-A monoclonal antibody into the solution containing the A antigen, adding an anti-B monoclonal antibody into the solution containing the B antigen, carrying out a fine immune flocculation reaction on the antigen and the antibody, uniformly mixing, and standing for 30 min; centrifuging at 10000rpm for 5min, and collecting precipitate;
(5) diffusing: mixing the collected precipitate with ultrapure water at a volume ratio of 1: 10, placing in 56 deg.C water bath for 10min, shaking, ultrafiltering, concentrating, and collecting membrane antigen. Obtaining pure erythrocyte membrane A antigen and B antigen;
the labeling method of colloidal gold is described in CN 103033632A;
a detection area is arranged on the reaction membrane 13, a first detection line 131, a second detection line 132 and a contrast line 133 are sequentially arranged on the detection area from the other end to one end of the reaction membrane 13, the first detection line 131 is coated with a solidified anti-A antibody, the second detection line 132 is coated with a solidified anti-B antibody, and the contrast line 133 is coated with a solidified anti-RBC antibody;
preferably, the above anti-a antibody and anti-B antibody are polyclonal antibodies, which are prepared by: according to the prior art, an anti-A/B antibody with an Fc fragment removed (which can be removed by an enzyme cutting method), namely a Fab fragment of the anti-A/B antibody, is used as an immunizing antigen to immunize a mouse for multiple times, and the titer and the specificity of a secondary antibody in serum are detected by blood drawing, wherein the titer is required to be not less than 1: 4096, and the specificity is 100%. Collecting the immunized mouse venous whole blood after reaching the standard, collecting serum, and purifying to obtain an anti-A antibody or an anti-B antibody;
preferably, the above anti-a antibody and anti-B antibody are monoclonal antibodies, which are prepared by: according to the prior art, an anti-A/B antibody with an Fc fragment removed (which can be removed by an enzyme cutting method), namely a Fab fragment of the anti-A/B antibody, is used as an immunizing antigen to immunize a mouse for multiple times, and spleen cells of the mouse are taken to be fused with tumor cells to obtain a hybridoma cell strain; and (3) screening hybridoma cell strains by the same method to obtain target cell strains, further culturing the target cell strains to obtain anti-A/B antibody specific secondary antibodies through secretion, or injecting the target cell strains into animals to obtain ascites, and purifying to obtain anti-A antibodies or anti-B antibodies.
As shown in fig. 2, the upper case 2 is disposed on the substrate 10 and covers the sample addition member 11, the antigen member 12, the reaction membrane 13, and the water absorption member 14, and the upper case 2 has a sample addition hole 21 corresponding to the sample addition member 11 and an observation window 22 corresponding to the detection area of the reaction membrane 13.
The utility model discloses an immunity chromatography detection technique, the anti A/B antibody of ABO blood group system in the qualitative detection human serum (thick liquid) sample. During detection, the blood group anti-A/B antibody in the sample can be combined with the labeled corresponding erythrocyte blood group A antigen and B antigen to form an antigen-antibody complex, and the complex moves forwards along the paper strip due to chromatography, and is combined with the pre-coated anti-A antibody and anti-B antibody to form an 'A/B antigen + anti-A/B antibody + anti-A/B antibody' sandwich after passing through the first detection line 131 and the second detection line 132 to form agglutination and color development. Free erythrocyte membrane anti-principle enriched color development at control line 133 was associated with pre-coated anti-RBC antibody. The negative specimen develops color only at the control line 133.
Preferably, the substrate 10 is made of a polyester plate; the material of the sample adding pad 11 is porous polymer material, has water conductivity and hydrophilicity, and can be glass fiber; the antigen pad 12 is made of porous polymer material; the reaction membrane 13 is a porous membrane, which can be a nitrocellulose membrane; the water absorption pad 14 is made of cotton pulp paper; the upper case 2 is made of plastic such as PS, ABS, PET, etc.
The utility model discloses a concrete application method as follows: 100uL serum (plasma) sample is dripped into the sample adding hole 21, the sample moves forwards along the paper strip under the action of chromatography, and the result is judged and read in the observation window 22 after waiting for 5-10 min. The interpretation method is as follows: the first detection line 131 is colored, an anti-A antibody exists in blood, the first detection line 131 is not colored, no anti-A antibody exists in blood, the second detection line 132 is colored, an anti-B antibody exists in blood, the second detection line 132 is not colored, no anti-B antibody exists in blood, and then whether the first detection line 131 and the second detection line 132 are colored or not is used for judging the corresponding blood type.
The above description is only a preferred embodiment of the present invention, and therefore the scope of the present invention should not be limited by this description, and all equivalent changes and modifications made within the scope and the specification of the present invention should be covered by the present invention.
Claims (6)
1. The utility model provides a reverse typing of people ABO blood group short-term test examination strip which characterized in that: the method comprises the following steps:
the test strip body comprises a substrate, and a sample adding pad, an antigen pad, a reaction film and a water absorbing pad which are arranged on the substrate, wherein the reaction film is arranged in the middle of the substrate, one end of the sample adding pad is contacted with the other end of the antigen pad, one end of the antigen pad is contacted with the other end of the reaction film, and one end of the reaction film is contacted with the other end of the water absorbing pad;
purified erythrocyte blood group A antigen and erythrocyte blood group B antigen marked by tracer particles are fixed on the antigen pad through physical adsorption; the reaction membrane is provided with a detection area, the detection area is sequentially provided with a first detection line, a second detection line and a contrast line from the other end to one end of the reaction membrane, the first detection line is coated with a solidified anti-A antibody, the second detection line is coated with a solidified anti-B antibody, and the contrast line is coated with a solidified anti-RBC antibody.
2. The human ABO blood group reverse typing rapid test strip as claimed in claim 1, wherein: the other end of the antigen pad is lapped on one end of the sample adding pad.
3. The human ABO blood group reverse typing rapid test strip as claimed in claim 1, wherein: one end of the antigen pad is lapped on the other end of the reaction membrane.
4. The human ABO blood group reverse typing rapid test strip as claimed in claim 1, wherein: the other end of the water absorption pad is lapped on one end of the reaction membrane.
5. The human ABO blood group reverse typing rapid test strip as claimed in claim 1, wherein: the tracer particles are colloidal gold.
6. The rapid test strip for human ABO blood group reverse typing according to any one of claims 1 to 5, wherein: the upper shell is arranged on the substrate and covers the sample adding pad, the antigen pad, the reaction membrane and the water absorption pad, and the upper shell is provided with a sample adding hole corresponding to the sample adding pad and an observation window corresponding to the detection zone of the reaction membrane.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202020368044.0U CN211905397U (en) | 2020-03-20 | 2020-03-20 | Human ABO blood type reverse-typing rapid detection test strip |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202020368044.0U CN211905397U (en) | 2020-03-20 | 2020-03-20 | Human ABO blood type reverse-typing rapid detection test strip |
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| CN211905397U true CN211905397U (en) | 2020-11-10 |
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| CN202020368044.0U Active CN211905397U (en) | 2020-03-20 | 2020-03-20 | Human ABO blood type reverse-typing rapid detection test strip |
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