CROSSMATCHING AND TRANSFUSION REACTION

Crossmatching (Compatibility Testing)

COLLECTION AND PREPARATION OF SAMPLES
Patient Identification Age of Specimen
• Very crucial; Common types of error • The freshest sample possible should be used for compatibility testing.
Collection • Specimen must be less than 3 days old if the patient has been transfused
• Serum is the preferred specimen for compatibility testing. Hemolysis or pregnant within the past 3 months.
should be avoided. Sample Storage
• Why serum not plasma? • The AABB requires that patient samples must be stored between 1-6⁰C
a. Plasma may cause small fibrin clots to form which may be for at least 7 days after transfusion.
difficult to distinguish from true agglutination.
b. Plasma may inactivate complement so that antibodies may not
be detected.
COMPATIBILITY TESTING PROTOCOLS
ABO Grouping Rh Typing
• Most critical pretransfusion serologic test • If the Rh type of the recipient cannot be determined and transfusion is
• If patient’s ABO group cannot be satisfactorily determined and immediate essential, Rh-negative blood should be given.
transfusion is essential, group O packed red cells should be utilized.
• Emergency cases (transfusion without knowing patient’s blood type): “O
negative” – Golden blood
Gel Test

Crossmatching
• Using AHG reagent (IAT) Purpose:
• Major X-match: Donor’s cells + Recipient’s serum a. Final check of ABO compatibility between patient and donor to prevent
• Minor X-match: Donor’s serum + Recipient’s cells transfusion reaction.
• 2-5% RCS b. Detects presence of antibody in patient’s serum that will react to donor’s
RBC that is not detected in antibody screen.
3 Phases of Crossmatching Reporting of Results
a. Immediate Spin in saline at RT – detects IgM • A compatible crossmatch is indicated by absence of agglutination
b. Thermophase / 37⁰C incubation for 30 mins. with enhancement and/or hemolysis at any stage of the crossmatch.
medium (albumin, LISS, PEG) – detects IgG • The absence of agglutination indicates that the patient has no
c. AHG Phase after washing incubated cells with saline. demonstrable antibodies with a specificity for any antigen on donor’s
RBC.
Check cells / Coombs control cells (IgG-sensitized cells) should be added to Ab Screen AC Major Xmatch Possible Problem
tubes that demonstrate no agglutination - - + • ABO/Rh typing error
- For results to be considered valid, agglutination must occur. • Donor unit w/ (+) DAT
• Patient w/ low incidence Ab
+ - + • Patient alloantibody
+ + + • Patient autoantibody
• Rouleaux
The Future of Compatibility Testing Blood Substitutes
• Red cell / Blood substitutes •
Substances that are able to carry oxygen in the absence of intact red
• Biochemical modification of non-O blood cells
• Galvanic biosensor – energy measured • For patients who are difficult to find compatible blood due to multiple
• Dipstick method of typing transfusions, it becomes challenging for them to obtain a compatible
• Dry plate method blood bag.
• For patients who can no longer receive blood from others.
• When there is a need to increase their blood volume.
BLOOD SUBSTITUTES (Cont’d)
Stroma-free Hb solutions / Hemoglobin-based Oxygen Carriers (HBOCs) Perfluorochemicals (PFCs)
Examples: PHP, PEG-Hb, Hemolink, Polyheme, HemAssist, Hemopure, Optro • Examples: Fluosol-DA-20, Oxygent
• Excellent gas (O2 and CO2) solvents

Dimasacat, Jeremiah S. | © Prof. Debbie Anne C. Rivera-De Leon

BLOOD BANKING
 Advantages of SFHS Disadvantages of SFHS Advantages of PFCs Disadvantages of PFCs
• Long shelf life • Short intravascular half-life • Biologic inertness • Adverse clinical effects
• Very stable • Possible toxicity • Lack of immunogenicity • High O2 affinity
• Not immunogenic • High O2 affinity • Easily synthesized • Retention in tissues
• No requirement for blood • High oncotic effect • For patients with low level of hemoglobin
typing procedures • Expensive; usually 2-3 doses intake
TRANSFUSION THERAPY
Lesion of Storage Autologous Transfusion
1. Decrease in glucose (due to cell consumption) It is a donation of blood by patients for transfusion to themselves in the future
2. Decrease pH (acidic) Emergency Transfusion
3. Build up of lactic acid • It is given to patients who are bleeding rapidly and uncontrollably.
4. Decrease ATP levels • Group O negative units should be used especially if the patient is a
5. Loss of red cell function woman of childbearing years.
6. Hemolysis
Massive Transfusion
7. Hyperkalemia
• Defined as the replacement of one or more blood volume/s within 24
8. Hyponatremia
hours or about 10 units of blood in an adult.
Adverse Conditions Associated with Massive Transfusion Neonatal Transfusion
• Citrate toxicity • Premature infants frequently require transfusion of small amounts of
• Hypocalcemia blood to replace blood drawn for laboratory tests.
• Hypothermia • Blood units less than 5 days old are preferred to lessen the risk of
• 2,3 DPG depletion hyperkalemia and to maximize the 2,3 DPG levels.
• Depletion of coagulation factors and platelets Exchange Transfusion
• Accumulation of biochemicals and microaggregates (To be discussed in HDN)

Transfusion Reactions
ACUTE TRANSFUSION REACTIONS
Immunologic
Acute / Immediate • Most severe and may be life threatening due to Febrile • Increase temperature of greater than 1⁰C after
Hemolytic ABO incompatibilities Nonhemolytic transfusion
Transfusion • The associated hemolysis is intravascular Transfusion • Mild immunonologic reactions that are caused by
Reaction (IHTR) • Mediators: IgM Abs (usually to ABO antigens), Reaction (FNHTR) the interaction of recipient antibodies against HLA
complement antigens on donor’s WBC and platelets
• S/S: fever, chills, hemoglobinuria, dyspnea, • Most common type of transfusion reactions
hypotension • Most common S/S: Fever and chills
• Most severe cases may result to DIC and renal • Management / Prevention: Use of leukocyte filters
failure during transfusion; Antipyretics
• Make sure that the patient has no fever before
blood transfusion.
• After releasing the blood bag, it will be warmed
using cloth to prevent it from getting cold. If the
patient suddenly develops a fever and
experiences chills during the ongoing
transfusion, you should immediately stop the
transfusion.
Allergic Transfusion • Second most common type of transfusion Anaphylactic • Mediator: Plasma, proteins, antibodies to IgA
Reaction reactions Transfusion (Anaphylactic reaction)
• IgE-mediated Reaction • Management / Prevention: Transfusion of IgA-
• S/S: Urticaria, Erythema, Hives, Itching, deficient components
Anaphylaxis
• Management / Prev79ention: Administration of
antihistamines before the transfusion
Noncardiogenic • Most consistent finding is Anti-leukocyte Abs in
Pulmonary Edema donor or patient plasma
(Aka Trali)
Non-Immunologic
Bacterial • Caused by the endotoxins produced by Gram- Transfusion • Good example of iatrogenic (physician-caused)
Contamination negative bacteria Associated transfusion reaction
• Mostly associated with cold growing Yersinia Circulatory • Common in patients with cardiac and pulmonary
enterocolitica, also with Pseudomonas spp. Overload (TACO) disease
(greenish) and Escherichia coli • May lead to congestive heart failure and
• The incidence of bacterial sepsis is highest with pulmonary edema
patients
• If you suspect bacterial contamination in the
blood bag, you need to return it to the

Dimasacat, Jeremiah S. | © Prof. Debbie Anne C. Rivera-De Leon

BLOOD BANKING
 bacteriology section for bacterial culturing. This
will help confirm if there are any growing
bacteria present as evidence of bacterial
contaminants.
DELAYED ADVERSE EFFECTS OF TRANSFUSION
Immunologic
Delayed Hemolytic • Characterized by the accelerated destruction of Transfusion • These reactions occur when immunologically
Transfusion transfused RBCs Associated Graft competent lymphocytes are transfused into an
Reaction (DHTR) • Most commonly associated with secondary Vs. Host Disease immunocompromised host
(anamnestic) response (TA-GVHD) • S/S: Fever, Liver problems, Rash, Diarrhea
• The associated hemolysis is generally • Management / Prevention: Transfusion of
extravascular irradiated blood components
• Mediators: IgG Abs to Rh, MNS, Kell, Kidd, and
Duffy antigens
Post-Transfusion • Rare transfusion reaction usually seen in older Graft-versus-Host Disease (GVHD)
Purpura female patients who have been sensitized to • Occurs when WBCs in transfused blood attack
platelet antigens, either by previous pregnancy the tissues of a transfusion recipient who has a
or transfusion. severely weakened immune.
• Characterized by severe thrombocytopenia 1 • Recipient’s Immune system doesn’t recognize the
week after the transfusion due to antibody to WBCs in the transfused blood as foreign. This
platelets specific antigens. allows them to survive and attack the recipient’s
body tissues.
• To prevent WBCs from causing GVHD, donated
blood can be treated with radiation before
transfusion.
Non-Immunologic
Transfusion-Induced • Iron deposition in vital organs seen in patients Transmission of • Hepatitis B, NANB Hepatitis (HCV), HIV, HTLV-1
Hemosiderosis who are thalassemic and with chronic Disease (oncogenic retrovirus that causes adult T cell
transfusion. leukemia), CMV, EBV
• Syphilis
o Although, transfusion of stored blood
has not been shown to transmit the
disease because spirochetes do not
survive at ref temp for 72 hours
• Hepatitis A
o Occurrence is very rare
o Infection by transfusion requires that
the donor has viremia (occurs briefly at
the same time of onset of acute
illness).

Dimasacat, Jeremiah S. | © Prof. Debbie Anne C. Rivera-De Leon

BLOOD BANKING