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Clinical Biochemistry is one of the aspects of laboratory medicinal techniques that is performed for the purpose of measuring the number of different types of chemicals that are present in the blood or in the urine, or in various fluids present within the human body. The results acquired through the tests are used in the cases of identification of the health problems, for the determination of the prognosis as well as finalizing the therapies for a patient. Blood transfusion, on the other hand, is the process of providing blood into the human body, which might have been lost due to surgical procedures, accidental injuries, or any other medical condition. The clinical assessment of the blood sample of the donor as well as the receiver is required to be clinically assessed to analyze the chemical properties of both the blood samples.
The information about the chemicals presented within both the blood samples provided an idea about the suitability between the blood provider and receiver (Chacham et al. 2019). The pieces of information about the blood serum testing and the chemicals present in them provide the chance of analyzing the compatibility of the blood samples with each other. The genes are considered sources of biological variation. The techniques of clinical assessment of the blood sample are performed include the Indirect Antiglobulin Technique (IAT) and the Enzyme panel technique. The principles of the assessment are to analyze the chemicals present within the blood serum of different samples and provide pieces of information about them before the process of blood transfusion.
The clinical assessment technique that will be used for analyzing the blood serum samples will be performed using the IAT techniques for analyzing the blood samples (Viberg et al. 2018). Therefore, various particulars of the IAT technique are discussed below.
Principles of IAT technique
This test is performed for the purpose of identification of the antibodies present within the red blood cells. The Antihuman Globulin (AHG) is used in this test for the purpose of detecting the In-vitro sensitivity of the red blood cells. In this procedure, the plasma of the patients is processed through incubation with the red cells that work as the reagents for the procedure. Moreover, in this process, an antigen phenotype is used which known in nature along with a solution is meant for potentiating. In the case of the presence of any antibodies of the red cell, it is meant to be bound with the reagent of the red cell along with the antigen that is corresponding.
There are various materials that are required for the purpose of analyzing the blood samples. These materials include
Low Ionic Strength Solution (LISS) is available from the Lorne Laboratories
Antihuman Globulin which is Polyspecific in nature
These materials are required for performing the clinical trial of the blood serums.
The procedure of analysis includes various steps such as
During this step all the preparations are made including the preparation of the reagents of the red cells, preparation of the water bath at 37degree Celsius, setting off the antigen identification panel, marking the tubes according to panel numbers and also by labeling the 11th tube to the tubes used for the “Pos control”.
Adding the Reagents
All the reagents including the samples of the blood of the patient, the reagents of the red cells are added and then they are added within the panel meant for the read cell. Moreover, 50µl of the LISS solution is also added to the panel. Then the temperature of the panel is set to be at 37 degrees Celsius and then the panel is left to incubate for 30 minutes.
Washing the cells
All the tubes are washed and centrifugation of each tube is performed at 1000g for a time period of 1 minute. The supernatant is decanted without making any disturbance to the button of the cell of each tube. Then the washing process is repeated for one more time including Saline. After this 100µl, AHG reagent is added to each of the tubes. After this, the tubes are centrifuged at 1000g following a rolling procedure in the light box. These are performed for agitating the tubes.
The samples are visually analyzed and it is actively marked as per their agglutination reactions.
Then after grading the agglutination reaction, the results are recorded against the cells of the provided antigram panel.
Some safety precautions that are required to be practiced while performing such tests include
Washing the reagents of the red cells, properly using lots of water as these reagents contain Sodium Azide, which forms an explosive compound by reacting with the copper in the pipes (Jackson et al. 2020).
Protective clothes must be used for protection while handling the materials such as the reagents of the red cells as well as the plasma due to the bio-hazardous nature of these materials.
Similar to the IAT technique there is also Direct Antiglobulin Testing (DAT) that is used for the clinical assessment of the blood samples before the blood transfusion process.
There are differences present in the procedure of DAT with that of IAT despite them being used for the same purpose of assessing blood serum samples. The Direct Antiglobulin Test is performed by the process of incorporation of the Antihuman Globulin (AHG) with the RBCs of the patients. On the other hand, in the case of the Indirect Antiglobulin Test, the plasma of the patients is used for testing the RBS after the addition of the Antihuman Globulin (AHG) is performed. Despite the difference in the processes, the RBC is agglutinated due to the presence of the Anti-RBC antibody, which might include either autoantibody as well as alloantibody. This agglutination occurs after the addition of the AHG is done (Bellairs et al. 2018).
The agglutination in the Direct Antiglobulin Test (DAT) occurs when the cross linking between an Antihuman globulin & Anti-RBC antibody occurs. The Anti-RBC antibodies are found to be bound with the antigen of the patient's RBCs.
On the other hand, in the case of the Indirect Antiglobulin Test (IAT), the process of agglutination is seen to be taking place when the cross-linking process takes place between the Antihuman Globulin with that of the Anti-RBC antibody present within the plasma of the patient. The RBCs for the test are added to this Anti-RBC antibody found in the plasma of the patient.
Despite these differences, the main purpose of these tests is to analyze the antigens and antibodies present within the two blood samples that include the patient as well as the donor samples before the process of blood sampling. This is very important for determining the compatibility of two blood samples before processing with the transfusion procedure.
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