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Antiglobulin assays, also known as Coombs tests, are considered commonly used diagnostic inventories used to detect immune infiltration and immunologic disorders. Depending on the technology and goals, two variants stand out. of antiglobulin tests: the direct antiglobulin test (DAT)/ direct Coombs’ Test (DCT) and Indirect antiglobulin Test (IAT). It is imperative to mention that direct antiglobulin This test is used to detect specificities such as anemia caused by a dysfunctional immune system. The human body can produce antibodies to detect and attack strange pathogens. In some cases, however, the immune system produces antibodies directed against red blood cells (or red blood cells in the body), and these can cause hemolytic anemia.
When should a direct antiglobulin test be performed?
If your doctor does not suspect whether or not you adhere to your red blood cells, direct antiglobulin test will help in these stories. Well – a known scenario where your doctor may recommend it! direct antiglobulin The test includes signs of post-transfusion hemolytic anemia and sometimes diagnosis in case the mother develops antibodies to fetal red blood cells. If undetected, the possibility of life-threatening disorders such as hemolytic disease of the newborn increases.
Hemolytic anemia is usually caused by activation of the immune system, leading to the formation of antibodies to the body’s own red blood cells, also known as autoimmune hemolytic anemia. The labeling has every opportunity to maintain
- Dyspnea
- Increased sense of lack of energy and lethargy.
- Dark urine color and yellow und.
- Low red blood cell count in blood tests.
- Cold hands and feet, especially hands and feet.
- Heart problems such as heart defects or irregular heartbeat.
How is the direct antiglobulin test performed?
How it works.
Direct antiglobulin This analysis helps demonstrate that the erythrocytes are covered with antibodies (the antibodies could be IgG or courtesy, or both). In the bloodstream, red blood cells have specific antigens that provide a place for antibodies. Any trigger or reaction that has the opportunity to activate these customs websites has every opportunity to initiate the signs of hemolytic anemia. This is believed to be the cause of HDN (hemolytic disease of the newborn), autoimmune hemolytic anemia, and transmissible reactions. Activation and platter have every opportunity to be triggered by certain medical drugs.
In this test, red blood cells are washed and incubated with human serum. This method ensures that the antibodies bind to the antigen on the erythrocytes. Next, the wash and anti-human globulin are added. If agglutination occurs after the addition of anti-human globulin (sometimes called Coombs-Lagen), this is considered a symptom of antibodies against the cells in vivo. It should be noted that this test can detect both IgG and IgM antibodies.
DAT Method
- Take the patient’s red blood cells from EDTA (Ethyl Endiamine Tactile Acid). This will pulverize the sample.
- Divide a drop of erythrocyte suspension (2%-5%) into one of four tubes individually and label the four tubes with the following labels: poly, control, anti-IGG, anti-C3.
- Rinse the saline solution with the saline solution and decant the physical solution by tilting the tubes upside down. Then clean the bottom of the tube with the left physical fabric and repeat this 3-4 times. Dry the tube with an absorbent cloth.
- Add 2 drops of poly-specific AHG to the “poly” tube and release 2 drops of 6% BSA into the “control” tube.
- Carefully shake to combine the contents of the tubes and form a rainfall or button that makes the red blood cells in each tube an agglutination test. Note the results.
- If poly-specific reagents show flattering results, cell coverage requires specific studies with anti-IGG anti-C3 reagent or antibody or complement or both.
- Divide two drops of anti-IGG AHG into “anti-IGG” tubes and two drops of anti-C3 AHG into “anti-C3” tubes. Shake to combine contents.
- All negatives should be incubated for 5 minutes for extensive coverage as the test needs to be very sensitive. To detect impotence in coverage, observe all negatives under a microscope.
- Use of a reddish upper tube to collect standards may result in false flattering results.
- If coagulation or cold occurs, this is due to debilitating autoantibodies against anti-I, including the effects of cold, this position is complemented in vitro.
- Avoid lavender tubes and bind calcium and magnesium ions.
- Avoid complement activation in vitro.
10. obtain correct information about the patient in case of positive
- diagnosis of a specific disease.
- Appropriate medications
- History of transfusion of red blood cells and plasma components
- Other studies showing destruction of red blood cells
To learn more about it and the IAT, watch the video below
How to interpret test results
We know that antibodies adhere to RBCs when they are flattered. The degree of positivity depends on the intensity of it and at the same time on the number of antibodies bound. However, this situation is not always associated with a positive result if the RBCs have already been destroyed. In direct antiglobulin test, antibodies are detected without identifying the cause, comparable to hemolytic anemia. To find the cause of this it is necessary to follow the circumstances of the disease, for example in the case of blood transfusions, reactions to medications, etc., a physical examination should be performed.
It is important to mention that only a small part of the population studies yield flattering results without the anxiety and symptoms of hemolytic anemia. If it is negative, it means that no antibodies adhere to the red blood cells. Therefore, other preconditions may be identified and investigated.
There are several stories in which it is flattering:
- Autoimmune hemolytic anemia (due to hyperactivity of the immune system against red blood cells)
- Development of antibodies on red blood cells, this situation is called “drug toxicity”. It is caused by some products such as NSAIDs like cephalosporod, levopod, dapsone, nitrofuran, ibuprofen, etc.
- When donated blood is attacked by the immune system (as part of a migratory response)
- Mothers and babies have different blood types.
- Leukemia, including acquired lymphocytic leukemia.
- An autoimmune disease called lupus.
- Monocleosis.
- Syphilis.
- Mycoplasma infestans.
Results are occasional. of direct antiglobulin Testing is not possible without disease or risk. In addition, the test may be interrupted by some products, some of them may cause flattering results without hemolytic anemia. Substances that may cause hemolytic anemia:
- Some antibiotics
- insulin
- Some specific blood pressure medications
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