COLLEGE OF HEALTH – HAIL Medical laboratory Dept.- Second term THIRD YEAR – Blood banking Compatibility test PRACTICE -7 INVESTIGATION OF A TRANSFUSION REACTION Adverse events related to transfusion can be acute (within 24 hours) or delayed (see Table below ). Transfusion laboratories should immediately be informed of a suspected transfusion reaction, being ideally placed to coordinate investigation, to communicate with clinicians and transfusion services, and to advise about appropriate choice of blood products for subsequent transfusions. Acute transfusion reactions are easier to attribute to the transfusion than delayed reactions, although, in patients who are already very ill, they can go undiagnosed. The symptoms and signs of acute transfusion reactions are similar regardless of the cause; treatment, and investigation of causes is simultaneous. It is easier to distinguish between the causes of delayed transfusion reactions, but it may be more difficult to recognise their relationship to the transfusion episode because of the delay in onset. The following scheme outlines the role of the laboratory in investigation and management of transfusion reactions, and a very useful algorithm can be found in the Handbook of Transfusion Medicine. Types of transfusion reaction Acute transfusion reactions Delayed transfusion reactions Acute haemolytic reaction Delayed haemolytic reaction Anaphylaxis Transfusion transmitted infection Bacterial contamination of blood product Transfusion-associated graft versus host disease Transfusion-associated acute lung injury Posttransfusion purpura Acute fluid overload Iron overload Allergic reaction Immunosuppression 1 Acute transfusion reactions Delayed transfusion reactions Febrile nonhaemolytic transfusion reaction — Acute Transfusion Reactions Acute life-threatening transfusion reactions can result from the following: 1. 2. 3. 4. 5. 6. Acute intravascular haemolysis as a result of ABO incompatibility Acute intravascular haemolysis can occur, although rarely as a result of other red cell antibodies that activate complement through to the membrane attack complex Severe extravascular haemolysis—this may happen where a strong antibody, which does not bind complement or only binds it to the C3 stage, is missed in pretransfusion testing and causes rapid extravascular clearance of incompatible transfused red cells. These reactions are usually less severe than those caused by ABO incompatibilities. Anaphylaxis and severe acute allergic reactions—these are more commonly associated with blood products containing large amounts of plasma where the recipient has been presensitised to an allergen in the donor plasma. Recipients with IgA deficiency can develop antibodies to IgA. Transfusion of an infected blood product—this is more common with platelets because they are stored at room temperature. If contamination is proven, the blood centre must be informed so that other components from the same donor can be traced. Transfusion-associated acute lung injury is an acute respiratory disorder, with one mechanism being passive transfer of antibodies in the donor unit that react with the recipient's own white blood cells, resulting in noncardiogenic interstitial pulmonary oedema. Although rare, the onset of acute transfusion reactions is usually very dramatic and the patient is acutely ill. Treatment is aimed at resuscitating the patient and elucidating the cause to try and prevent any further incidents ( Table below ). In addition, there are unpleasant but not life-threatening reactions that may occur during transfusion. They include the following: 2 Allergic reactions—a mild urticaria or itching caused by a reaction to plasma proteins in the donor unit Immediate investigations in the case of an acute transfusion reaction Check for haemolysis Perform visual examination of patient's plasma and urine (plasma and urine haemoglobin can be checked but this is not essential). Blood film will show spherocytosis, red cell fragmentation. Bilirubin and lactate dehydrogenase (LDH) levels will be raised. Check for incompatibility Check the documentation and the patient's identity. Repeat ABO group of patient pretransfusion and posttransfusion and of the donor unit(s). Screen the patient for red cell antibodies pretransfusion and posttransfusion. Repeat crossmatch with pretransfusion and posttransfusion samples. Direct antiglobulin test (DAT) on patient. Eluate from patient's red cells if DAT is positive. Check for disseminated intravascular coagulation Perform blood count and film, coagulation screen, and fibrin degradation products (or Ddimers). Check for renal function Check blood urea, creatinine, and electrolytes. Check for bacterial infections Take blood cultures from the patient and donor unit including immediate gram stain. Immunological investigations Check immunoglobulin A (IgA) levels and anti-IgA antibodies. Febrile non-haemolytic transfusion reactions—recipient's antibodies that react to donor white cells and cause an increase in temperature of no more that 1°C; alternatively, cytokines released from white cells in the donor units can cause a similar reaction. These conditions usually settle on slowing the transfusion and administration of antipyretics and antihistamines. They do not require detailed investigation. Acute Intravascular Haemolysis 3 Transfused red cells react with the patient's own anti-A or anti-B, and the red cells are destroyed in the circulation, causing collapse, renal failure, and disseminated intravascular coagulation. Transfusion of ABO-incompatible cells usually results from an identification error. This can occur at point of blood sampling and labelling (wrong blood in tube), laboratory testing (technical error), blood unit labelling (administrative error), and collection from the blood refrigerator or inadequate bedside checking. If red cells are mistakenly transfused to the wrong patient, there is approximately a 1 in 3 chance that ABO incompatibility will occur. The reaction is most severe if group A blood is transfused to a patient who is group O, and only a few millilitres of red cells are required to cause this reaction. Prompt action in recognising this acute emergency and stopping the transfusion may lead to a better outcome because the severity depends on the volume of blood transfused. If an acute transfusion reaction is suspected, the laboratory must be informed immediately and the unit of blood and giving set must be returned to the laboratory with blood and urine samples from the patient ( Table 20.10 ). Documentation Check Patient identification, the compatibility form, and the compatibility label of the blood unit should be checked again at the bedside. Any discrepancies must be notified to the transfusion laboratory immediately. If the wrong blood has been administered, the units intended for that patient must be withdrawn from issue to prevent another parallel error occurring with another patient who may have the same or a similar name. Serological Investigations Serological investigations have a twofold purpose: (a) to check for any laboratory errors in the pretransfusion sample group and compatibility check and (b) to repeat the group and compatibility tests with the posttransfusion sample to see if the pretransfusion sample was from the correct patient. Reactions in liquid-phase tests should be read microscopically to detect any mixed-field reaction. Tests for Haemolysis Because not all acute transfusion reactions are the result of haemolysis, haematological and biochemical tests as well as visual inspection of the plasma/serum and urine are required (see Chapter 9 ). Further tests may be required to manage the resuscitation of the patient and direct the use of blood products to treat disseminated intravascular coagulation. Microbiological Tests If the cause of the acute transfusion reaction is still unclear, blood cultures should be taken from the unit and the patient. Blood centres issue guidance for the investigation of potentially contaminated units. Delayed Haemolytic Transfusion Reaction 4 A delayed haemolytic transfusion reaction occurs when the recipient has been immunised to a red cell antigen by a previous transfusion or during pregnancy but the antibody is present at low or undetectable levels. A secondary immune response is mounted to the incompatible antigen that has been transfused. The IgG- and/or complement-coated red cells are destroyed in the spleen and liver. Kidd antibodies are often implicated in delayed transfusion reactions because they are difficult to detect, often displaying a dosage effect, fall rapidly to undetectable levels, and are frequently present in combinations of antibodies. Haematological Investigation The following suggest a delayed haemolytic transfusion reaction: Haemoglobin concentration falls more rapidly than would be expected after a red cell transfusion Increase in haemoglobin concentration is less than expected for the number of units transfused Blood film shows spherocytosis Positive direct antiglobulin test Unconjugated bilirubin raised Serological Investigation It is desirable to have the pretransfusion sample available to test in addition to a posttransfusion sample, but this is not always possible because of the delay between the time of the transfusion and the investigation. It has been recommended by some that plasma/serum samples are saved on all patients who are transfused, but this is not always practical. Unless the reaction is acute, the units transfused will not be available for retesting. In the United Kingdom, the phenotype of each unit is provided by the National Blood Service, and this information can help in the investigation of a delayed transfusion reaction. The following tests should be carried out, preferably using different or more sensitive techniques: 1. 2. 3. 4. Confirm the ABO and D group of the patient on a pretransfusion and posttransfusion sample. Perform a direct antiglobulin test on the patient's pretransfusion and posttransfusion washed red cells. In the event of a positive direct antiglobulin test, elution of the antibody may aid identification or confirm specificities in cases of non-ABO incompatibility. Repeat the crossmatch, if possible, using pretransfusion and posttransfusion samples. Screen the pretransfusion and posttransfusion samples for red cell antibodies and identify any antibodies. The immediate posttransfusion sample may have no detectable red cell antibodies, al-though they may be eluted from the patient's red cells if the direct antiglobulin test is positive. It is also possible to have a delayed 5 haemolytic transfusion reaction with a negative direct antiglobulin test because the antibody-coated red cells have been removed from the circulation. If the immediate posttransfusion investigation is inconclusive, repeat the tests 10 days later to allow antibody levels to increase 6