Preparation method of newborn ABO hemolytic disease detection simulation specimen for teaching
By preparing simulated specimens, the problem of difficulty in obtaining specimens for neonatal ABO hemolytic disease testing was solved, and the simulation of intuitive operations in teaching was achieved to meet teaching needs.
Patent Information
- Application Number
- CN202410263942.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-03-08
- Publication Date
- 2025-09-09
AI Technical Summary
In the teaching of transfusion medicine, it is difficult to obtain specimens for neonatal ABO hemolytic disease testing, which makes teaching difficult. Existing methods mainly rely on lectures and video teaching, and lack intuitive operation experience.
A method for preparing simulated specimens includes absorbing IgM antibodies and IgG antibodies, mixing healthy human plasma and packed red blood cells, and preserving the mixture after centrifugation to simulate the detection process of neonatal ABO hemolytic disease and prepare type A or type B simulated specimens.
It provides simulated specimens that can be used for teaching, helping students understand and master experimental operations, solving the problem of difficulty in obtaining specimens, and meeting teaching needs.
Smart Images
Figure CN120612855A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the field of transfusion medicine teaching, and particularly relates to a method for preparing a simulated specimen for detecting neonatal ABO hemolytic disease in transfusion medicine teaching. Background Art
[0002] Hemolytic disease of the newborn (HDN) is caused by red blood cell incompatibility between mother and fetus. The mother's immune system is sensitized by red blood cells from the fetus, producing corresponding IgG blood type antibodies. These antibodies enter the fetal blood circulation through the placenta, causing increased destruction of alloimmune newborn (fetal) red blood cells and resulting in extravascular hemolysis, leading to anemia, edema, and hepatosplenomegaly. It is one of the most common acute hemolytic diseases in pediatrics. Severe cases can endanger the child's life or be accompanied by sequelae such as kernicterus.
[0003] Common blood type-related antibodies are mostly IgM antibodies and IgG antibodies. The blood type antibodies that cause HDN are almost all IgG antibodies, which can pass through the placental barrier and are strongly reactive at 37°C. They are called warm antibodies, while IgM blood type antibodies are strongly reactive at 4°C and are called cold antibodies.
[0004] In my country, HDN (ABO hemolytic disease of the newborn) caused by ABO blood incompatibility is the most common. Diagnosis is primarily based on the newborn's clinical manifestations combined with the results of three serological hemolysis tests, the latter of which are crucial for diagnosis. These tests, including the direct antiglobulin test (DAT), the cytokine release test, and the ionization test, are essential for a definitive diagnosis of ABO HDN. The DAT detects whether the child's red blood cells are sensitized by maternal IgG antibodies. The cytokine release test detects the presence of maternal IgG antibodies corresponding to the child's own blood type in the patient's plasma. The ionization test detects the presence of maternal IgG antibodies corresponding to the child's own blood type bound to the surface of the child's red blood cells.
[0005] In the teaching of transfusion medicine, the detection of neonatal ABO hemolytic disease is an important experimental operation project. However, since neonatal blood samples are not easy to obtain, it is difficult to conduct experimental teaching through intuitive and practical operations. Teaching can only be carried out through lectures and operation videos, which is not conducive to the development of teaching work and it is difficult to achieve teaching objectives. Summary of the Invention
[0006] The purpose of the present invention is to provide a method for preparing a simulated specimen for detecting ABO hemolytic disease of newborns in the teaching of transfusion medicine. The method of the present invention can prepare a simulated specimen for detecting ABO hemolytic disease of newborns for use in teaching, which is conducive to the development of teaching work.
[0007] In order to achieve the above technical objectives, the present invention adopts the following technical solution: a method for preparing a simulated specimen for the detection of neonatal ABO hemolytic disease for teaching, characterized in that it comprises the following steps: (1) Absorption of IgM anti-A, anti-B, and anti-AB: Take healthy human type O plasma or serum, type A packed red blood cells, type B packed red blood cells, and type AB packed red blood cells in a volume ratio of 20:2:2:1, transfer them into a centrifuge tube, mix them thoroughly, and then install the lid. After the absorption reaction, obtain a centrifuge tube containing the mixture; (2) Separation of absorbed plasma or serum: Place the centrifuge tube containing the mixture in a centrifuge. After centrifugation, draw out the upper layer of plasma or serum for later use. (3) Absorption of IgG anti-A or anti-B by type A or type B packed red blood cells: take type A or type B packed red blood cells of healthy individuals and the plasma or serum prepared in step (2) at a volume ratio of 1:3, transfer them into another centrifuge tube, mix them thoroughly, and tighten the lid. The mixture obtained after the absorption reaction is the corresponding simulated specimen for type A or type B neonatal ABO hemolytic disease detection; (4) Specimen preservation: The simulated specimen for neonatal ABO hemolytic disease test prepared in step (3) is stored in a refrigerator at 2-8°C for future use.
[0008] The type A packed red blood cells, type B packed red blood cells, and type AB packed red blood cells are the lower layer red blood cells obtained by centrifuging whole blood of the corresponding blood types at 3000 rpm for 3 minutes.
[0009] Preferably, in step (1), 5 mL of healthy human type O plasma or serum, 0.5 mL of type A packed red blood cells, 0.5 mL of type B packed red blood cells, and 0.25 mL of type AB packed red blood cells are respectively transferred into centrifuge tubes, mixed thoroughly, and then covered.
[0010] In step (1), the absorption reaction needs to be placed in a 4°C environment for 10 minutes, during which the centrifuge tube needs to be inverted and mixed once.
[0011] Preferably, the plasma or serum prepared in step (2) needs to be subjected to blood typing reverse typing to verify whether the IgM anti-A, anti-B and anti-AB are completely absorbed. If no agglutination reaction occurs during reverse typing, the absorption is complete. If an agglutination reaction occurs, the absorption is incomplete and it needs to be re-prepared according to steps (1) and (2).
[0012] The upper layer of plasma or serum prepared in step (2) needs to be tested for IgG anti-A or anti-B using the anti-human globulin method. If an agglutination reaction occurs, the preparation is successful; if no agglutination reaction occurs, the preparation fails and needs to be re-prepared according to steps (1) and (2).
[0013] Preferably, in step (3), the absorption reaction needs to be incubated at 37°C for 1 hour, during which the centrifuge tube needs to be inverted and mixed twice.
[0014] In step (3), 1 mL of type A or type B packed red blood cells from a healthy person and 3 mL of the plasma or serum prepared in step (2) are transferred into another centrifuge tube, mixed thoroughly, and then the tube is covered.
[0015] The present invention has the beneficial effect of preparing a simulated specimen using the method. The specimen can be used in the testing of neonatal ABO hemolytic disease, an experimental component of transfusion medicine laboratory teaching. The use of simulated specimens facilitates students' learning and understanding of experimental techniques, addresses the difficulty in obtaining specimens for neonatal ABO hemolytic disease testing, and meets the needs of transfusion medicine laboratory teaching. BRIEF DESCRIPTION OF THE DRAWINGS
[0016] Figure 1 It is a flow chart of the preparation method of the present invention.
[0017] Figure 2 This is the ABO blood type test result of the simulated specimen in the present invention.
[0018] Figure 3 This is the test result of ABO hemolytic disease of newborn in the simulated specimen in the present invention. DETAILED DESCRIPTION
[0019] The technical solutions in the embodiments of the present invention will be described clearly and completely below in conjunction with the accompanying drawings of the present invention.
[0020] like Figure 1 As shown, the present invention provides a method for preparing a simulated specimen for detecting neonatal ABO hemolytic disease in blood transfusion medicine teaching, comprising the following steps: Absorption of IgM anti-A, anti-B, and anti-AB: Take 5 mL of healthy human type O plasma, 0.5 mL of type A packed red blood cells, 0.5 mL of type B packed red blood cells, and 0.25 mL of type AB packed red blood cells, respectively. Transfer them into centrifuge tubes, mix thoroughly, install the cap, tighten it, and incubate at 4°C for 10 minutes. During this time, invert the tube once to mix thoroughly. Obtain a centrifuge tube containing the mixture.
[0021] (2) Separation of absorbed plasma: Place the centrifuge tube containing the mixture in a centrifuge and centrifuge at 3000 rpm for 3 minutes. Aspirate the upper layer of plasma or serum and keep it for later use. (3) Absorption of IgG anti-A or anti-B by type A or type B packed red blood cells: 1 mL of type A or type B packed red blood cells from a healthy individual and 3 mL of plasma prepared in step (2) were transferred into centrifuge tubes, mixed thoroughly, and the caps were tightened. The tubes were incubated at 37°C for 1 h.
[0022] (4) Specimen preservation: The mixture prepared in step (3) is the simulated specimen for the detection of type A or type B neonatal ABO hemolytic disease, which should be stored in a refrigerator at 2-8°C for future use.
[0023] In the above preparation method, the type A packed red blood cells, type B packed red blood cells, and type AB packed red blood cells are the lower layer of red blood cells obtained by centrifuging whole blood of the corresponding blood type at 3000 rpm for 3 minutes. The healthy human type O plasma can also be replaced with healthy human type O serum to achieve the preparation of simulated specimens.
[0024] The simulated specimen prepared by the present invention is used to detect neonatal ABO hemolytic disease (taking type A as an example), including positive typing of neonatal ABO blood type, direct antiglobulin test, free test, radiation test and control experiment. The specific steps are as follows: (1) Simulated sample ABO blood type test: Take the ABO and RhD blood type test cards, add 50uL of the prepared simulated sample 1% red blood cell suspension into the positive typing A and B holes respectively, and use a dedicated blood type serology centrifuge to centrifuge at 900rpm for 2 minutes and 1500rpm for 3 minutes. The results are as follows: Figure 2 As shown, the ABO blood type of the simulated specimen is type A; (2) Add 50uL of the prepared 1% red blood cell suspension of the simulated specimen to the low-ion anti-human globulin card and mark it with direct antibody. Incubate it in a dedicated incubator at 37°C for 15 minutes and centrifuge it in a dedicated centrifuge for 8 minutes. The results are as follows: Figure 3 As shown, the direct anti-human globulin test of the simulated specimen was positive; (3) Detect IgG blood type antibodies in the free test, emission test and control test (final washing solution). Take a low-ion anti-human globulin card and mark the first 3 wells, the middle 3 wells and the last 3 wells as the free test, emission test and control test respectively. Add 50uL of 1% A, B and O reagent red blood cells to each of the 3 wells in each group and mark them. Add 25uL of plasma to each of the first 3 wells and mark them as the free test, add 25uL of emission solution to each of the middle 3 wells and mark them as the emission test, add 25uL of the last washing solution to each of the last 3 wells and mark them as the control test. Use a special incubator to incubate at 37℃ for 15 minutes and a special centrifuge to centrifuge for 8 minutes. The results are as follows: Figure 3 As shown, it is consistent with the detection pattern of neonatal type A hemolytic disease.
[0025] The specimens prepared using this method are simulated specimens that can be used in the neonatal ABO hemolytic disease testing component of transfusion medicine laboratory teaching. The use of simulated specimens facilitates students' learning and understanding of experimental techniques, addresses the difficulty in obtaining specimens for neonatal ABO hemolytic disease testing, and meets the needs of transfusion medicine laboratory teaching.
[0026] Any details not described in detail herein are well known to those skilled in the art. Finally, it should be noted that the above embodiments are intended only to illustrate the technical solutions of the present invention and are not intended to limit the present invention. Although the present invention has been described in detail with reference to preferred embodiments, it should be understood by those skilled in the art that the technical solutions of the present invention may be modified or replaced with equivalents without departing from the spirit and scope of the technical solutions of the present invention, and such modifications or equivalents should be encompassed by the claims of the present invention.
Claims
1. A method for preparing a simulated specimen for testing neonatal ABO hemolytic disease for teaching purposes, characterized by: The following steps are involved: (1) Absorption of IgM anti-A, anti-B, and anti-AB: Take healthy human type O plasma or serum, type A packed red blood cells, type B packed red blood cells, and type AB packed red blood cells in a volume ratio of 20:2:2:1, transfer them into a centrifuge tube, mix them thoroughly, and then install the lid. After the absorption reaction, obtain a centrifuge tube containing the mixture; (2) Separation of absorbed plasma or serum: Place the centrifuge tube containing the mixture in a centrifuge. After centrifugation, draw out the upper layer of plasma or serum for later use. (3) Absorption of IgG anti-A or anti-B by type A or type B packed red blood cells: take type A or type B packed red blood cells of healthy individuals and the plasma or serum prepared in step (2) at a volume ratio of 1:3, transfer them into another centrifuge tube, mix them thoroughly, and tighten the lid. The mixture obtained after the absorption reaction is the corresponding simulated specimen for type A or type B neonatal ABO hemolytic disease detection; (4) Specimen preservation: The simulated specimen for neonatal ABO hemolytic disease test prepared in step (3) is stored in a refrigerator at 2-8°C for future use.
2. The method for preparing a simulated specimen for detecting ABO hemolytic disease of the newborn for teaching purposes according to claim 1, wherein: The type A packed red blood cells, type B packed red blood cells, and type AB packed red blood cells are the lower layer red blood cells obtained by centrifuging whole blood of the corresponding blood types at 3000 rpm for 3 minutes.
3. The method for preparing a simulated specimen for detecting ABO hemolytic disease of the newborn for teaching purposes according to claim 1, wherein: In step (1), 5 mL of healthy human type O plasma or serum, 0.5 mL of type A packed red blood cells, 0.5 mL of type B packed red blood cells, and 0.25 mL of type AB packed red blood cells were transferred into centrifuge tubes, mixed thoroughly, and then covered.
4. The method for preparing a simulated specimen for detecting ABO hemolytic disease of the newborn for teaching purposes according to claim 1, wherein: In step (1), the absorption reaction needs to be placed in a 4°C environment for 10 minutes, during which the centrifuge tube needs to be inverted and mixed once.
5. The method for preparing a simulated specimen for detecting ABO hemolytic disease of the newborn for teaching purposes according to claim 1, wherein: The plasma or serum prepared in step (2) needs to be subjected to reverse typing to verify whether the IgM anti-A, anti-B and anti-AB are completely absorbed. If no agglutination reaction occurs after reverse typing, the absorption is complete. If an agglutination reaction occurs, the absorption is incomplete and it needs to be re-prepared according to steps (1) and (2).
6. The method for preparing a simulated specimen for detecting ABO hemolytic disease of the newborn for teaching purposes according to claim 1, characterized in that: The upper layer of plasma or serum prepared in step (2) needs to be tested for IgG anti-A or anti-B using the anti-human globulin method. If an agglutination reaction occurs, the preparation is successful; if no agglutination reaction occurs, the preparation fails and needs to be re-prepared according to steps (1) and (2).
7. The method for preparing a simulated specimen for detecting ABO hemolytic disease of the newborn for teaching purposes according to claim 1, characterized in that: In step (3), the absorption reaction needs to be incubated at 37°C for 1 hour, during which the centrifuge tube needs to be inverted and mixed twice.
8. The method for preparing a simulated specimen for detecting ABO hemolytic disease of the newborn for teaching purposes according to any one of claims 1 to 7, characterized in that: In step (3), 1 mL of type A or type B packed red blood cells from a healthy person and 3 mL of the plasma or serum prepared in step (2) are transferred into another centrifuge tube, mixed thoroughly, and the cap is tightened.