CN113092793A - Method and system for detecting Rh blood group antigen and application thereof - Google Patents
Method and system for detecting Rh blood group antigen and application thereof Download PDFInfo
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- 239000000427 antigen Substances 0.000 title claims abstract description 72
- 102000036639 antigens Human genes 0.000 title claims abstract description 72
- 108091007433 antigens Proteins 0.000 title claims abstract description 72
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Abstract
The invention relates to a method and a system for detecting Rh blood group antigen and application thereof, belonging to the technical field of biological detection. The invention provides a positive typing method for detecting Rh blood group antigen based on solid phase adsorption, which comprises the steps of adding red blood cells to be typed and Rh blood group antibody into a container coated with other species of second antibody for centrifugation, observing whether the red blood cells to be typed are adsorbed at the bottom of the container coated with other species of second antibody, if the red blood cells to be typed are adsorbed at the bottom of the container coated with other species of second antibody, the Rh blood group antigen corresponding to the red blood cells to be typed and the Rh blood group antibody is positive, and if the red blood cells to be typed are not adsorbed at the bottom of the container coated with other species of second antibody, the Rh blood group antigen corresponding to the red blood cells to be typed and the Rh blood group antibody is negative; the method has the advantages of high sensitivity, strong specificity and low detection cost, is easy for automatic operation, and can avoid human errors in the operation process.
Description
Technical Field
The invention relates to a method and a system for detecting Rh blood group antigen and application thereof, belonging to the field of biological detection.
Background
The erythrocyte blood type is a classical human genetic marker and plays an important role in the fields of blood transfusion, paternity test, anthropology research, forensic physical evidence inspection and the like.
At present, 43 blood group systems of red blood cells are known, wherein the meaning of ABO blood group system in clinical blood transfusion is the most important, and Rh blood group system is the second place. The Rh blood group system is one of the most complex blood group systems in the human erythroid system, and has 18 phenotypes, 46 blood group antigens, including D antigen, C antigen, E antigen, etc., wherein the D antigen is particularly important in hemolytic newborn caused by clinical blood transfusion and blood group incompatibility of mother and infant, and therefore, D antigen positive is often called Rh positive, and D antigen negative is called Rh negative.
The Rh blood group incompatible blood transfused into the blood recipient stimulates the immune system of the blood recipient to produce alloantibody which may cause extravascular hemolysis when the blood recipient transfuses again, so the D antigen in the Rh blood group system is listed as the item to be checked in the clinical transfusion technical Specification for testing before transfusion. In recent years, hemolytic reaction and cross matching difficulty caused by other Rh blood group antigens than the D antigen have been frequently reported, and neonatal hemolysis caused by the Rh blood group system has also frequently occurred. Therefore, the detection of Rh blood group system before transfusion of a transfusion subject, especially for some patients who need repeated transfusions, and the detection of Rh blood group system for women of childbearing age, pregnant women, and fetuses, is of great importance for preventing post-transfusion hemolysis and neonatal hemolysis caused by Rh blood group system.
Blood type typing can be divided into positive typing and negative typing. The positive typing is to identify the presence or absence of corresponding blood group antigens on erythrocytes with blood group antibodies, and the negative typing is to identify the presence or absence of corresponding blood group antibodies in the serum (plasma) of a subject with erythrocytes with antigens. Currently, the general Rh blood typing method uses the principle that blood group antibodies and erythrocytes undergo agglutination reaction in vitro, and the common methods include a test tube method, a microplate method, and a microcolumn gel method (the test tube method, the microplate method, and the microcolumn gel method are all described in "national clinical test operating procedures").
However, these methods all have drawbacks, such as:
the test tube method requires a manual mixing step, which is time-consuming and labor-consuming, and the test tube needs to be shaken when the test tube method is used for judging the agglutination degree, so that the detection result is easily influenced by the force, different persons with the same sample may generate different judgment results, and especially for some weak positive samples, manual errors are easily generated;
the result stability is influenced by the sample adding amount of the antibody and the erythrocyte and the difference between antibody batches in the microplate method;
the micro-column gel method is easy to deform and generate bubbles in the transportation process, and the temperature also influences the size of the gel molecular sieve, so that the stability and reliability of the final detection result are influenced.
Disclosure of Invention
In order to solve the above problems, the present invention provides a method for detecting Rh blood group antigen, comprising a typing step; the parting step is as follows: adding the red blood cells to be typed and the Rh blood group antibody into a container coated with other species of second antibodies for centrifugation, observing whether the red blood cells to be typed are adsorbed at the bottom of the container coated with other species of second antibodies after the centrifugation is finished, if the red blood cells to be typed are adsorbed at the bottom of the container coated with other species of second antibodies, the Rh blood group antigen corresponding to the red blood cells to be typed and the Rh blood group antibody is positive, and if the red blood cells to be typed are not adsorbed at the bottom of the container coated with other species of second antibodies, the Rh blood group antigen corresponding to the red blood cells to be typed and the Rh blood group antibody is negative.
In one embodiment of the invention, the Rh blood group antibody comprises Rh blood group antibodies of various species origin; the Rh blood group antibodies of various species sources comprise IgM antibodies and IgG antibodies of various species sources, which are used for resisting human erythrocyte Rh system blood group antigens.
In one embodiment of the invention, when the species is human, the IgM antibody comprises at least one of an anti-D antibody, an anti-C antibody, an anti-E antibody and an anti-E antibody.
In one embodiment of the invention, the other species secondary antibody comprises a secondary antibody of the other species against the corresponding Rh blood group antibody; the secondary antibodies of the other species against the corresponding Rh blood group antibodies comprise at least one of IgM antibodies of the other species against the corresponding Rh blood group antibodies and IgG antibodies of the other species against the corresponding Rh blood group antibodies. For example, when the Rh blood group antibody is a human Rh blood group antibody, the other species secondary antibody comprises at least one of a goat anti-human secondary antibody comprising at least one of a goat anti-human IgM antibody and a goat anti-human IgG antibody, and a mouse anti-human secondary antibody comprising at least one of a mouse anti-human IgM antibody and a mouse anti-human IgG antibody.
In one embodiment of the present invention, the typing step is:
adding the red blood cells to be typed and the RhD blood type antibody into a container coated with other species of second antibodies for centrifugation, observing whether the red blood cells to be typed are adsorbed at the bottom of the container coated with other species of second antibodies after the centrifugation is finished, if the red blood cells to be typed are adsorbed at the bottom of the container coated with other species of second antibodies, the red blood cells to be typed are Rh D positive, and if the red blood cells to be typed are not adsorbed at the bottom of the container coated with other species of second antibodies, the red blood cells to be typed are Rh D negative;
and/or adding the red blood cells to be typed and the RhC blood group antibody into a container coated with a second antibody of other species for centrifugation, observing whether the red blood cells to be typed are adsorbed at the bottom of the container coated with the second antibody of other species after the centrifugation is finished, if the red blood cells to be typed are adsorbed at the bottom of the container coated with the second antibody of other species, the red blood cells to be typed are Rh C positive, and if the red blood cells to be typed are not adsorbed at the bottom of the container coated with the second antibody of other species, the red blood cells to be typed are Rh C negative;
and/or adding the red blood cells to be typed and the Rhc blood group antibody into a container coated with a second antibody of other species for centrifugation, observing whether the red blood cells to be typed are adsorbed at the bottom of the container coated with the second antibody of other species after the centrifugation is finished, if the red blood cells to be typed are adsorbed at the bottom of the container coated with the second antibody of other species, the red blood cells to be typed are Rh c positive, and if the red blood cells to be typed are not adsorbed at the bottom of the container coated with the second antibody of other species, the red blood cells to be typed are Rh c negative;
and/or adding the red blood cells to be typed and the RhE blood group antibodies into a container coated with other species secondary antibodies for centrifugation, observing whether the red blood cells to be typed are adsorbed at the bottom of the container coated with other species secondary antibodies or not after the centrifugation is finished, if the red blood cells to be typed are adsorbed at the bottom of the container coated with other species secondary antibodies, the red blood cells to be typed are Rh E positive, and if the red blood cells to be typed are not adsorbed at the bottom of the container coated with other species secondary antibodies, the red blood cells to be typed are Rh E negative;
and/or adding the red blood cells to be typed and Rhe blood group antibodies into a container coated with other species secondary antibodies for centrifugation, observing whether the red blood cells to be typed are adsorbed at the bottom of the container coated with other species secondary antibodies or not after the centrifugation is finished, if the red blood cells to be typed are adsorbed at the bottom of the container coated with other species secondary antibodies, the red blood cells to be typed are Rh e positive, and if the red blood cells to be typed are not adsorbed at the bottom of the container coated with other species secondary antibodies, the red blood cells to be typed are Rh e negative.
In one embodiment of the present invention, the rotation speed of the centrifugation is 90 to 360g and the time is 1 to 30 min.
In one embodiment of the invention, the rotation speed of the centrifugation is 190-360 g, and the time is 1-3 min.
In one embodiment of the present invention, the typing step further comprises a dilution step; the dilution step is as follows: diluting the red blood cells to be classified to the concentration of 0.3-1%.
In one embodiment of the present invention, the diluting step is: the red blood cells to be typed are diluted to a concentration of 0.3 to 1% by Liss (low ionic strength solution) or normal saline.
In one embodiment of the present invention, the diluting step is: washing the red blood cells to be typed for 3-5 times by using normal saline, and then diluting the red blood cells to be typed to the concentration of 0.3-1% by using Liss or normal saline.
In one embodiment of the invention, the amount of the red blood cells to be typed in the container coated with the second antibody of other species is 10-100 μ L.
In one embodiment of the present invention, the container coated with the second antibody of the other species is prepared by: adding second antibodies of other species into the container, and coating for 8-16 h at 2-8 ℃ to obtain a coated container; and adding a buffer solution containing BSA (bovine serum albumin) and Tween20 (Tween-20) into the coated container, and blocking at 37 ℃ for 1-6 h to obtain a container coated with a second antibody of other species.
In one embodiment of the invention, the buffer solution is a carbonate buffer solution with a pH of 8.9-9.8; the volume concentration of the BSA in the buffer solution is 0.1-5%; the volume concentration of the Tween20 in the buffer solution is 0.02-1%.
In one embodiment of the present invention, the method for preparing the container coated with the second antibody of the other species comprises the following steps:
coating: diluting the second antibody of other species to the concentration of 0.5-10 mu g/mL by using a carbonate buffer solution with the pH of 8.9-9.8 to obtain a diluent; adding the diluent into a container, and coating for 8-16 h at 2-8 ℃ to obtain a coated container;
a first plate washing step: adding PBS buffer solution with pH of 6.5-7.8 into the coated container for washing, and repeatedly washing for 3-5 times to obtain a washed container;
and (3) sealing: adding a carbonate buffer solution with the pH of 8.9-9.8, wherein the carbonate buffer solution contains BSA (bovine serum albumin) with the volume concentration of 0.1-5% and Tween20 with the volume concentration of 0.02-1%, and sealing at 37 ℃ for 1-6 h to obtain a sealed container;
and a second plate washing step: and adding PBS buffer solution with pH of 6.5-7.8 into the sealed container for washing, and repeating the washing for 3-5 times to obtain a container coated with the second antibody of other species.
In one embodiment of the invention, the container is a microplate.
In one embodiment of the present invention, the microplate is a U-shaped microplate.
The invention also provides a system for detecting by using the method, wherein the system comprises a container coated with a second antibody of other species and Rh blood group antibodies.
In one embodiment of the invention, the container is a microplate.
In one embodiment of the present invention, the microplate is a U-shaped microplate.
In one embodiment of the present invention, the container coated with the second antibody of the other species is prepared by: adding second antibodies of other species into the container, and coating for 8-16 h at 2-8 ℃ to obtain a coated container; and adding a buffer solution containing BSA and Tween20 into the coated container, and blocking at 37 ℃ for 1-6 h to obtain a container coated with a second antibody of other species.
In one embodiment of the invention, the buffer solution is a carbonate buffer solution with a pH of 8.9-9.8; the volume concentration of the BSA in the buffer solution is 0.1-5%; the volume concentration of the Tween20 in the buffer solution is 0.02-1%.
In one embodiment of the present invention, the method for preparing the container coated with the second antibody of the other species comprises the following steps:
coating: diluting the second antibody of other species to the concentration of 0.5-10 mu g/mL by using a carbonate buffer solution with the pH of 8.9-9.8 to obtain a diluent; adding the diluent into a container, and coating for 8-16 h at 2-8 ℃ to obtain a coated container;
a first plate washing step: adding PBS buffer solution with pH of 6.5-7.8 into the coated container for washing, and repeatedly washing for 3-5 times to obtain a washed container;
and (3) sealing: adding a carbonate buffer solution with the pH of 8.9-9.8, wherein the carbonate buffer solution contains BSA (bovine serum albumin) with the volume concentration of 0.1-5% and Tween20 with the volume concentration of 0.02-1%, and sealing at 37 ℃ for 1-6 h to obtain a sealed container;
and a second plate washing step: and adding PBS buffer solution with pH of 6.5-7.8 into the sealed container for washing, and repeating the washing for 3-5 times to obtain a container coated with the second antibody of other species.
The invention also provides the application of the method or the system in the detection of the Rh blood group antigen.
The technical scheme of the invention has the following advantages:
the invention provides a positive typing method for detecting Rh blood group antigen based on solid phase adsorption, which comprises the steps of adding red blood cells to be typed and Rh blood group antibody into a container coated with other species of second antibody for centrifugation, observing whether the red blood cells to be typed are adsorbed at the bottom of the container coated with other species of second antibody after the centrifugation is finished, if the red blood cells to be typed are adsorbed at the bottom of the container coated with other species of second antibody, the Rh blood group antigen corresponding to the red blood cells to be typed and the Rh blood group antibody is positive, and if the red blood cells to be typed are not adsorbed at the bottom of the container coated with other species of second antibody, the Rh blood group antigen corresponding to the red blood cells to be typed and the Rh blood group antibody is negative; compared with a test tube method, the method avoids manual operation, has the advantages of easy automation operation and high stability, has less consumption of coating antibodies, ensures that sample red blood cells are also trace after dilution, has clear and readable reaction results, has the advantages of high sensitivity and strong specificity, and has the advantages of getting rid of dependence on gel with higher price compared with a micro-column gel method, identifying all Rh blood group antigens of the sample red blood cells by 5 micropores and trace antibodies and the sample red blood cells, and has the advantage of low cost.
Drawings
FIG. 1: a detection schematic diagram of a method for detecting Rh blood group antigen. In fig. 1, from left to right: the kit comprises a container coated with other species of second antibodies, a container coated with other species of second antibodies added with red blood cells to be classified and Rh blood group antibodies, a container coated with other species of second antibodies with red blood cells adsorbed, and a container coated with other species of second antibodies with red blood cells not adsorbed.
FIG. 2: a detection result chart of a method for detecting Rh blood group antigen. In fig. 2, the positive results are that red blood cells are adsorbed by the second antibody of other species at the bottom of the container under the mediation of the Rh blood group antibody, and the positive results are divided into 4+, 3+, 2+ and 1+ according to the degree of adsorption; the negative result is that red blood cells cannot be adsorbed by other species of secondary antibodies at the bottom of the container mediated by Rh blood group antibodies, thereby forming a circular cell button at the bottom of the container.
FIG. 3: rh blood group typing experiment result chart (anti-C \ C \ E \ E antibody).
FIGS. 4 to 5: graph of Rh blood grouping test results (test tube method and microcolumn gel method are used as controls).
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: method for detecting Rh blood group antigen (Rh anti-D)
This example provides a method for detecting Rh blood group antigen (the detection principle is shown in fig. 1, and the possible detection result is shown in fig. 2), the method includes the following steps:
(1) coating: diluting the mouse anti-human IgM antibody to a concentration of 1 mug/mL by using a carbonate buffer solution with a pH of 9.6 to obtain a diluent; adding the diluent into a U-shaped microporous plate with the addition amount of 100 mu L per hole, and then coating for 12h at 4 ℃ to obtain a coated microporous plate;
(2) a first plate washing step: adding 250 mu L of PBS buffer solution with pH 7.2 into the coated microplate for washing according to the addition amount of each hole, repeating the washing for 5 times, wherein the residual liquid in the microplate needs to be sucked and dried in each washing to obtain a washed microplate A;
(3) and (3) sealing: adding a carbonate buffer solution with pH 9.6 and containing BSA with the volume concentration of 1% and Tween20 with the volume concentration of 0.02% into the washed microplate A by the adding amount of 250 mu L per well, and then sealing for 2h at 37 ℃ to obtain a sealed microplate;
(4) adding 250 mu L of PBS buffer solution with pH 7.2 into the sealed microplate for washing according to the addition amount of each hole, repeating the washing for 5 times, wherein the residual liquid in the microplate is required to be sucked and dried in each washing, and obtaining a washed microplate B;
(5) diluting: washing with physiological saline for typing red blood cells for 3 times, and diluting with Liss for typing red blood cells respectively until the concentration is 0.5% to obtain red blood cell suspension to be typed;
(6) and (3) detection: adding the suspension of the red blood cells to be typed into the washed microplate B respectively according to the addition amount of 75 microliter per hole, simultaneously adding the humanized IgM anti-D antibody into the washed microplate B according to the addition amount of 25 microliter per hole, and centrifuging for 1min at the rotating speed of 190g to obtain a centrifuged microplate; observing the centrifuged microporous plate, if the red blood cells to be classified are adsorbed at the bottom of the container coated with the mouse anti-human IgM antibody, the red blood cells to be classified are Rh D positive, and if the red blood cells to be classified are not adsorbed at the bottom of the container coated with the mouse anti-human IgM antibody, the red blood cells to be classified are Rh D negative.
Example 2: method for detecting Rh blood group antigen (Rh anti-D)
This example provides a method for detecting Rh blood group antigen (the detection principle is shown in fig. 1, and the possible detection result is shown in fig. 2), the method includes the following steps:
(1) coating: diluting the mouse anti-human IgM antibody to a concentration of 0.5 mug/mL by using a carbonate buffer solution with a pH of 8.9 to obtain a diluent; adding the diluent into a U-shaped microporous plate by the addition amount of 120 mu L per hole, and then coating for 8h at 2 ℃ to obtain a coated microporous plate;
(2) a first plate washing step: adding 200 mu L of PBS buffer solution with pH 6.5 into the coated microplate for washing according to the addition amount of 200 mu L of each hole, repeating the washing for 3 times, wherein the residual liquid in the microplate needs to be sucked and dried in each washing to obtain a washed microplate A;
(3) and (3) sealing: adding a carbonate buffer solution with pH 8.9 and containing BSA with the volume concentration of 0.1% and Tween20 with the volume concentration of 0.02% into the washed microplate A by the addition amount of 200 mu L per well, and then sealing for 1h at 37 ℃ to obtain a sealed microplate;
(4) adding 200 mu L of PBS buffer solution with pH 6.5 into the sealed microporous plate for washing, repeating the washing for 3 times, wherein the residual liquid in the microporous plate needs to be sucked dry in each washing to obtain a washed microporous plate B;
(5) diluting: washing the red blood cells to be typed with normal saline for 3 times, and then respectively diluting the red blood cells to be typed with Liss until the concentration is 0.3 percent to obtain red blood cell suspension to be typed;
(6) and (3) detection: adding the suspension of the red blood cells to be typed into the washed microplate B respectively according to the addition amount of 90 microliter per hole, simultaneously adding the humanized IgM anti-D antibody into the washed microplate B according to the addition amount of 10 microliter per hole, and centrifuging for 1min at the rotating speed of 190g to obtain a centrifuged microplate; observing the centrifuged microporous plate, if the red blood cells to be classified are adsorbed at the bottom of the container coated with the mouse anti-human IgM antibody, the red blood cells to be classified are Rh D positive, and if the red blood cells to be classified are not adsorbed at the bottom of the container coated with the mouse anti-human IgM antibody, the red blood cells to be classified are Rh D negative.
Example 3: method for detecting Rh blood group antigen (Rh anti-D)
This example provides a method for detecting Rh blood group antigen (the detection principle is shown in fig. 1, and the possible detection result is shown in fig. 2), the method includes the following steps:
(1) coating: diluting the mouse anti-human IgM antibody to a concentration of 10 mug/mL by using a carbonate buffer solution with a pH of 9.8 to obtain a diluent; adding the diluent into a U-shaped microporous plate with the addition amount of 100 mu L per hole, and then coating for 16h at 8 ℃ to obtain a coated microporous plate;
(2) a first plate washing step: adding 300 mu L of PBS buffer solution with pH 7.8 into the coated microplate for washing according to the addition amount of 300 mu L of each hole, repeating the washing for 5 times, and sucking the residual liquid in the microplate for each washing to obtain a washed microplate A;
(3) and (3) sealing: adding a carbonate buffer solution with pH 9.8 and containing BSA with the volume concentration of 5% and Tween20 with the volume concentration of 1% into the washed microplate A by the addition amount of 300 mu L per well, and then sealing for 2h at 37 ℃ to obtain a sealed microplate;
(4) adding 300 mu L of PBS buffer solution with pH 7.8 into the sealed microplate for washing according to the addition amount of 300 mu L of each hole, repeating the washing for 5 times, wherein the residual liquid in the microplate is required to be sucked and dried in each washing, and obtaining a washed microplate B;
(5) diluting: washing the red blood cells to be typed with normal saline for 5 times, and then respectively diluting the red blood cells to be typed with Liss until the concentration is 1 percent to obtain red blood cell suspension to be typed;
(6) and (3) detection: adding the suspension of the red blood cells to be typed into the washed microplate B respectively according to the addition amount of 80 microliter per hole, simultaneously adding the humanized IgM anti-D antibody into the washed microplate B according to the addition amount of 20 microliter per hole, and centrifuging for 3min at the rotating speed of 360g to obtain a centrifuged microplate; observing the centrifuged microporous plate, if the red blood cells to be classified are adsorbed at the bottom of the container coated with the mouse anti-human IgM antibody, the red blood cells to be classified are Rh D positive, and if the red blood cells to be classified are not adsorbed at the bottom of the container coated with the mouse anti-human IgM antibody, the red blood cells to be classified are Rh D negative.
Example 4: method for detecting Rh blood group antigen (Rh anti-D)
This example provides a method for detecting Rh blood group antigen (the detection principle is shown in fig. 1, and the possible detection result is shown in fig. 2), the method includes the following steps:
(1) coating: diluting the mouse anti-human IgM antibody to a concentration of 5 mug/mL by using a carbonate buffer solution with a pH of 9.6 to obtain a diluent; adding the diluent into a U-shaped microporous plate with the addition amount of 100 mu L per hole, and then coating for 12h at 4 ℃ to obtain a coated microporous plate;
(2) a first plate washing step: adding 250 mu L of PBS buffer solution with pH 7.2 into the coated microplate for washing according to the addition amount of each hole, repeating the washing for 5 times, wherein the residual liquid in the microplate needs to be sucked and dried in each washing to obtain a washed microplate A;
(3) and (3) sealing: adding a carbonate buffer solution with pH 9.6 and containing BSA with the volume concentration of 1% and Tween20 with the volume concentration of 0.02% into the washed microplate A by the adding amount of 250 mu L per well, and then sealing for 2h at 37 ℃ to obtain a sealed microplate;
(4) adding 250 mu L of PBS buffer solution with pH 7.2 into the sealed microplate for washing according to the addition amount of each hole, repeating the washing for 5 times, wherein the residual liquid in the microplate is required to be sucked and dried in each washing, and obtaining a washed microplate B;
(5) diluting: washing with physiological saline for typing red blood cells for 3 times, and diluting with Liss for typing red blood cells respectively until the concentration is 0.5% to obtain red blood cell suspension to be typed;
(6) and (3) detection: adding the red blood cell suspension to be typed into the washed microplate B respectively according to the addition amount of 95 microliter per hole, simultaneously adding the human IgM anti-D antibody into the washed microplate B according to the addition amount of 5 microliter per hole, and centrifuging for 30min at the rotating speed of 90g to obtain a centrifuged microplate; observing the centrifuged microporous plate, if the red blood cells to be classified are adsorbed at the bottom of the container coated with the mouse anti-human IgM antibody, the red blood cells to be classified are Rh D positive, and if the red blood cells to be classified are not adsorbed at the bottom of the container coated with the mouse anti-human IgM antibody, the red blood cells to be classified are Rh D negative.
Examples 5 to 8: method for detecting Rh blood group antigen (Rh anti-E)
The embodiment provides a method for detecting Rh blood group antigens (the detection principle is shown in figure 1, and possible detection results are shown in figure 2), which is characterized in that human IgM anti-D antibodies are replaced by human IgM anti-E antibodies on the basis of embodiments 1-4; observing the centrifuged microporous plate, if the red blood cells to be classified are adsorbed at the bottom of the container coated with the mouse anti-human IgM antibody, the red blood cells to be classified are Rh E positive, and if the red blood cells to be classified are not adsorbed at the bottom of the container coated with the mouse anti-human IgM antibody, the red blood cells to be classified are Rh E negative.
Examples 9 to 12: method for detecting Rh blood group antigen (Rh anti e)
The embodiment provides a method for detecting Rh blood group antigens (the detection principle is shown in figure 1, and possible detection results are shown in figure 2), which is characterized in that human IgM anti-D antibodies are replaced by human IgM anti-e antibodies on the basis of embodiments 1-4; observing the centrifuged microporous plate, if the red blood cells to be typed are adsorbed at the bottom of the container coated with the mouse anti-human IgM antibody, the red blood cells to be typed are Rh e positive, and if the red blood cells to be typed are not adsorbed at the bottom of the container coated with the mouse anti-human IgM antibody, the red blood cells to be typed are Rh e negative.
Examples 13 to 16: method for detecting Rh blood group antigen (Rh anti-C)
The embodiment provides a method for detecting Rh blood group antigen (the detection principle is shown in figure 1, and the possible detection result is shown in figure 2), which is characterized in that on the basis of the embodiments 1-4, an anti-human IgM anti-D antibody is replaced by a human IgM anti-C antibody; observing the centrifuged microporous plate, if the red blood cells to be typed are adsorbed at the bottom of the container coated with the mouse anti-human IgM antibody, the red blood cells to be typed are Rh C positive, and if the red blood cells to be typed are not adsorbed at the bottom of the container coated with the mouse anti-human IgM antibody, the red blood cells to be typed are Rh C negative.
Examples 17 to 20: method for detecting Rh blood group antigen (Rh anti-c)
The embodiment provides a method for detecting Rh blood group antigens (the detection principle is shown in figure 1, and possible detection results are shown in figure 2), which is characterized in that human IgM anti-D antibodies are replaced by human IgM anti-c antibodies on the basis of embodiments 1-4; observing the centrifuged microporous plate, if the red blood cells to be typed are adsorbed at the bottom of the container coated with the mouse anti-human IgM antibody, the red blood cells to be typed are Rh c positive, and if the red blood cells to be typed are not adsorbed at the bottom of the container coated with the mouse anti-human IgM antibody, the red blood cells to be typed are Rh c negative.
Experimental example 1: rh blood group antigen typing test (reaction of anti-C, anti-C, anti-E, anti-E antibodies with erythrocytes as an example)
1.1 Experimental materials
1.1.1 Primary reagents and consumables
Reagent: human anti-D IgM antibody (titer 256, available from Millipore corporation), human anti-C IgM antibody (titer 32, available from Millipore corporation), human anti-E IgM antibody (titer 8, available from Millipore corporation), murine anti-human IgM secondary antibody (available from thermoliser corporation), carbonate buffer at pH 9.6 (available from Sigma corporation), PBS buffer at pH 7.2 (available from Sigma corporation), Tween20 (available from Sigma corporation), BSA (available from thermoliser corporation), Liss (available from Jiangsu Leibo pharmaceutical Biotechnology GmbH), two red cell samples of unknown Rh blood group antigens.
Consumable material: test tube centrifuge, microplate centrifuge, full-automatic microplate operating system, 10 ~ 1000 uL application of sample rifle, 96 orifice plates can be dismantled to U type (purchase from NUNC Maxisorp company).
1.2Rh blood group antigen typing experimental method
Rh blood group antigen typing experiment Using the method for detecting Rh blood group antigens of examples 5, 9, 13 and 17, the following steps were included:
1.2.1 coating: diluting a mouse anti-human IgM second antibody to the concentration of 1 mu g/mL by using a carbonate buffer solution with the pH value of 9.6 to obtain a diluent; adding the diluent into a U-shaped detachable 96 micro-porous plate according to the addition amount of 100 mu L per hole, and then coating for 12h at 4 ℃ to obtain a coated micro-porous plate;
1.2.2 first plate washing step: adding 250 mu L of PBS buffer solution with pH 7.2 into the coated microplate for washing according to the addition amount of each hole, repeating the washing for 5 times, wherein the residual liquid in the microplate needs to be sucked and dried in each washing to obtain a washed microplate A;
1.2.3 blocking: adding a carbonate buffer solution with pH 9.6 and containing BSA with the volume concentration of 1% and Tween20 with the volume concentration of 0.02% into the washed microplate A by the adding amount of 250 mu L per well, and then sealing for 2h at 37 ℃ to obtain a sealed microplate;
1.2.4 adding 250 microliter of PBS buffer solution with pH 7.2 into the sealed microporous plate for washing, repeating the washing for 5 times, wherein the residual liquid in the microporous plate needs to be sucked and dried in each washing to obtain a washed microporous plate B;
1.2.5 dilution: respectively washing two red blood cell samples of unknown Rh blood group antigens for 3 times by using normal saline, and respectively diluting the two red blood cell samples of unknown Rh blood group antigens to the concentration of 0.3% by using Liss to obtain two to-be-typed red blood cell suspensions;
1.2.6 detection: adding two parts of erythrocyte suspension to be determined into the washed microporous plate B respectively according to the addition amount of 75 microliter per hole, simultaneously adding human anti-C IgM antibody, human anti-C IgM antibody, human anti-E IgM antibody and human anti-E IgM antibody into the washed microporous plate B respectively according to the addition amount of 25 microliter per hole, and centrifuging for 1min at the rotating speed of 190g to obtain a centrifuged microporous plate; observation of centrifuged microplates
1.3 results of Rh blood group antigen typing experiment
As shown in fig. 3, 4 wells of each sample, the first 4 wells from top to bottom, are the detection results of the first sample, and the detection results are: c positive, C negative, E positive; the detection result of the second sample is 4 holes from top to bottom, and the detection result is as follows: c-negative, C-positive, E-negative.
Experimental example 2: accuracy experiment of Rh blood group antigen typing method
2.1 materials of the experiment
See experimental example 1.
2.2 accuracy test method of Rh blood group antigen typing method
The accuracy experimental method of the Rh blood group antigen typing method uses a test tube method and a microcolumn gel method (the test tube method and the microcolumn gel method are recorded in national clinical test operating procedures) as a control, adopts the same detection reagent and method of the experimental example 1, and has the difference that two red cell samples of unknown Rh blood group antigens are replaced by the other two red cell samples of unknown Rh blood group antigens; the detection results are observed and shown in FIGS. 4-5.
2.3 accuracy test results of Rh blood group antigen typing method
As shown in FIGS. 4 to 5, the results of Rh blood group antigen typing of red blood cell samples of unknown Rh blood group antigens using the detection reagent and method of example 1 were consistent with those of the test tube method and the microcolumn gel method (wherein the typing results of the red blood cell samples in FIG. 4 are C + C-E-E +, and the typing results of the red blood cell samples in FIG. 5 are C-C + E + E-). As can be seen, the results of Rh blood group antigen typing of erythrocytes using the detection reagent and method of example 1 were accurate.
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. A method for detecting Rh blood group antigens, comprising a typing step; the parting step is as follows: adding the red blood cells to be typed and the Rh blood group antibody into a container coated with other species of second antibodies for centrifugation, observing whether the red blood cells to be typed are adsorbed at the bottom of the container coated with other species of second antibodies after the centrifugation is finished, if the red blood cells to be typed are adsorbed at the bottom of the container coated with other species of second antibodies, the Rh blood group antigen corresponding to the red blood cells to be typed and the Rh blood group antibody is positive, and if the red blood cells to be typed are not adsorbed at the bottom of the container coated with other species of second antibodies, the Rh blood group antigen corresponding to the red blood cells to be typed and the Rh blood group antibody is negative.
2. The method of claim 1, wherein the Rh blood group antibody comprises Rh blood group antibodies from various species; when the species is human, the Rh blood group antibody comprises at least one of an anti-D antibody, an anti-C antibody, an anti-E antibody, and an anti-E antibody.
3. The method of claim 2, wherein the second antibody of another species comprises a second antibody of another species against the corresponding Rh blood group antibody; the secondary antibodies of the other species against the corresponding Rh blood group antibodies comprise at least one of IgM antibodies and IgG antibodies.
4. A method for detecting Rh blood group antigens according to claim 2 or 3, wherein the typing step is:
adding the red blood cells to be typed and the RhD blood type antibody into a container coated with other species of second antibodies for centrifugation, observing whether the red blood cells to be typed are adsorbed at the bottom of the container coated with other species of second antibodies after the centrifugation is finished, if the red blood cells to be typed are adsorbed at the bottom of the container coated with other species of second antibodies, the red blood cells to be typed are Rh D positive, and if the red blood cells to be typed are not adsorbed at the bottom of the container coated with other species of second antibodies, the red blood cells to be typed are Rh D negative;
and/or adding the red blood cells to be typed and the RhC blood group antibody into a container coated with a second antibody of other species for centrifugation, observing whether the red blood cells to be typed are adsorbed at the bottom of the container coated with the second antibody of other species after the centrifugation is finished, if the red blood cells to be typed are adsorbed at the bottom of the container coated with the second antibody of other species, the red blood cells to be typed are Rh C positive, and if the red blood cells to be typed are not adsorbed at the bottom of the container coated with the second antibody of other species, the red blood cells to be typed are Rh C negative;
and/or adding the red blood cells to be typed and the Rhc blood group antibody into a container coated with a second antibody of other species for centrifugation, observing whether the red blood cells to be typed are adsorbed at the bottom of the container coated with the second antibody of other species after the centrifugation is finished, if the red blood cells to be typed are adsorbed at the bottom of the container coated with the second antibody of other species, the red blood cells to be typed are Rh c positive, and if the red blood cells to be typed are not adsorbed at the bottom of the container coated with the second antibody of other species, the red blood cells to be typed are Rh c negative;
and/or adding the red blood cells to be typed and the RhE blood group antibodies into a container coated with other species secondary antibodies for centrifugation, observing whether the red blood cells to be typed are adsorbed at the bottom of the container coated with other species secondary antibodies or not after the centrifugation is finished, if the red blood cells to be typed are adsorbed at the bottom of the container coated with other species secondary antibodies, the red blood cells to be typed are Rh E positive, and if the red blood cells to be typed are not adsorbed at the bottom of the container coated with other species secondary antibodies, the red blood cells to be typed are Rh E negative;
and/or adding the red blood cells to be typed and Rhe blood group antibodies into a container coated with other species secondary antibodies for centrifugation, observing whether the red blood cells to be typed are adsorbed at the bottom of the container coated with other species secondary antibodies or not after the centrifugation is finished, if the red blood cells to be typed are adsorbed at the bottom of the container coated with other species secondary antibodies, the red blood cells to be typed are Rh e positive, and if the red blood cells to be typed are not adsorbed at the bottom of the container coated with other species secondary antibodies, the red blood cells to be typed are Rh e negative.
5. The method for detecting Rh blood group antigen according to any one of claims 1 to 4, wherein the rotation speed of the centrifugation is 90 to 360g and the time is 1 to 30 min.
6. The method for detecting Rh blood group antigen according to any one of claims 1 to 5, further comprising a dilution step before the typing step; the dilution step is as follows: diluting the red blood cells to be classified to the concentration of 0.3-1%.
7. The system for detecting Rh blood group antigen according to any one of claims 1 to 6, wherein the system comprises a container coated with a second antibody of another species and Rh blood group antibodies.
8. The system of claim 7, wherein the container is a microwell plate.
9. The system of claim 7 or 8, wherein the container coated with the second antibody of the other species is prepared by: adding second antibodies of other species into the container, and coating for 8-16 h at 2-8 ℃ to obtain a coated container; and adding a buffer solution containing BSA and Tween20 into the coated container, and blocking at 37 ℃ for 1-6 h to obtain a container coated with a second antibody of other species.
10. Use of a method of detecting Rh blood group antigens according to any one of claims 1 to 6 or a system according to any one of claims 7 to 9 for detecting Rh blood group antigens.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115753652A (en) * | 2022-11-03 | 2023-03-07 | 江苏力博医药生物技术股份有限公司 | Method for quantitatively detecting RhD antigen by using erythrocyte endogenous peroxidase |
Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1904618A (en) * | 2006-07-31 | 2007-01-31 | 潘干华 | Method of implementing erythrocyte blood group antigen detection on haemocyte analysis instrument |
| US20070218515A1 (en) * | 2006-03-14 | 2007-09-20 | Nina Zhou | Method and kits for the determination of the antigens on the red blood cells and antibodies of serum |
| CN101109755A (en) * | 2007-08-20 | 2008-01-23 | 陕西省血液中心 | Reagent kit used for screening irregular antibody in blood serum and preparing method thereof |
| US20080318798A1 (en) * | 2005-03-24 | 2008-12-25 | Colin Campbell | Antigen Detection |
| US20110076698A1 (en) * | 2008-06-10 | 2011-03-31 | Intec Products, Inc. (Xiamen) | method of quickly determining human abo/rh/mn blood type and a test kit thereof |
| CN102175877A (en) * | 2010-12-31 | 2011-09-07 | 周胜利 | Method for trace, fast and accurate detection of human erythrocyte antigen and antibodies in serum |
| CN103926415A (en) * | 2014-03-24 | 2014-07-16 | 上海市血液中心 | System for combined and rapid detection of blood type antibody in human blood |
| US20170059591A1 (en) * | 2014-02-21 | 2017-03-02 | Qbd (Qs-Ip) Limited | Red blood cell detection |
| CN107643409A (en) * | 2017-09-19 | 2018-01-30 | 汪德清 | A kind of blood group antigens chip and its application in red blood cell accident antibody test |
| CN108680756A (en) * | 2018-05-21 | 2018-10-19 | 中国科学院苏州生物医学工程技术研究所 | A kind of incomplete antibody detection kit and detection method |
| CN110174518A (en) * | 2019-04-16 | 2019-08-27 | 骆宏 | Sorting type erythrocyte blood type irregular antibody detection kit based on solid phase agglutination technology and preparation method thereof |
| CN111638373A (en) * | 2020-06-08 | 2020-09-08 | 长春博德生物技术有限责任公司 | Kit for ABO blood group positive typing and Rh blood group detection and preparation method and detection method thereof |
-
2021
- 2021-04-16 CN CN202110413003.8A patent/CN113092793A/en active Pending
Patent Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080318798A1 (en) * | 2005-03-24 | 2008-12-25 | Colin Campbell | Antigen Detection |
| US20070218515A1 (en) * | 2006-03-14 | 2007-09-20 | Nina Zhou | Method and kits for the determination of the antigens on the red blood cells and antibodies of serum |
| CN1904618A (en) * | 2006-07-31 | 2007-01-31 | 潘干华 | Method of implementing erythrocyte blood group antigen detection on haemocyte analysis instrument |
| CN101109755A (en) * | 2007-08-20 | 2008-01-23 | 陕西省血液中心 | Reagent kit used for screening irregular antibody in blood serum and preparing method thereof |
| US20110076698A1 (en) * | 2008-06-10 | 2011-03-31 | Intec Products, Inc. (Xiamen) | method of quickly determining human abo/rh/mn blood type and a test kit thereof |
| CN102175877A (en) * | 2010-12-31 | 2011-09-07 | 周胜利 | Method for trace, fast and accurate detection of human erythrocyte antigen and antibodies in serum |
| US20170059591A1 (en) * | 2014-02-21 | 2017-03-02 | Qbd (Qs-Ip) Limited | Red blood cell detection |
| CN103926415A (en) * | 2014-03-24 | 2014-07-16 | 上海市血液中心 | System for combined and rapid detection of blood type antibody in human blood |
| CN107643409A (en) * | 2017-09-19 | 2018-01-30 | 汪德清 | A kind of blood group antigens chip and its application in red blood cell accident antibody test |
| CN108680756A (en) * | 2018-05-21 | 2018-10-19 | 中国科学院苏州生物医学工程技术研究所 | A kind of incomplete antibody detection kit and detection method |
| CN110174518A (en) * | 2019-04-16 | 2019-08-27 | 骆宏 | Sorting type erythrocyte blood type irregular antibody detection kit based on solid phase agglutination technology and preparation method thereof |
| CN111638373A (en) * | 2020-06-08 | 2020-09-08 | 长春博德生物技术有限责任公司 | Kit for ABO blood group positive typing and Rh blood group detection and preparation method and detection method thereof |
Non-Patent Citations (1)
| Title |
|---|
| 胡丽华: "《人类红细胞血型学实用理论与实验技术》", vol. 1, 31 August 2004, 中国医药科技出版社, pages: 255 - 257 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115753652A (en) * | 2022-11-03 | 2023-03-07 | 江苏力博医药生物技术股份有限公司 | Method for quantitatively detecting RhD antigen by using erythrocyte endogenous peroxidase |
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