CROSSMATCHING Albano, Astorga, Dumangon, Fregil, Gutierrez, Loque 4AMT Pretransfusion Compatibility Testing

a.k.a. Compatibility Testing Series of testing procedures and processes with the ultimate objective of ensuring the best possible result (safety) of a blood transfusion The primary objective of the crossmatch test is to detect the presence of antibodies in the recipients serum that could destroy transfused RBCs.

Phases of Crossmatching
Immediate Saline Spin Phase Also known as Protein / Albumin / Room Temperature phase Accomplished by mixing the patients serum with donors RBCs (Major) and the donors serum with patients RBCs (Minor) then centrifuge it immediately Absence of agglutination or hemolysis indicates compatibility Detects: Incompatibility in ABO system Incompatibility due to cold antibodies Prozoning anti-Rh antibodies are detected in a serum albumin mixture on immediate centrifugation

Crossmatch testing / Crossmatching

Traditionally meant the testing of the patients serum with the donor RBCs including an antiglobulin phase or simply an immediate spin phase to confirm ABO compatibility Two main functions of serologic crossmatching: It serves as a final check of ABO compatibility between donor and patient. It may detect the presence of an antibody in the patients serum that will react with antigens on the donor RBCs but that was not detected in Ab screening because the corresponding Ag was lacking from the screening cells. Thermo Phase

Also known as Incubation phase In this phase, the tubes showing no agglutination in the immediate spin phase will be incubated for 30 minutes at 37C water bath (10 mins if LISS is used instead of 22% BSA) then centrifuge No agglutination or hemolysis indicates compatibility Detects Incompatibility due to presence of low titered anti-Rh Certain Rh antibodies (anti-C, E and some D)

It is divided into 2 parts: MAJOR Crossmatch Patients serum + Donors red cells (PS-DR)

MINOR Crossmatch Donors serum+ Patients red cells (DS-PR)

CROSSMATCHING Albano, Astorga, Dumangon, Fregil, Gutierrez, Loque 4AMT

Antihuman Globulin Phase Also known as Coombs phase For greater sensitivity, AHG containing both anti-IgG and anti-complement may be used blend of rabbit anti-IgG and murine monoclonal anti-complement In this phase, the cells of tubes showing no agglutination in the previous phase are washed with NSS thrice. Next, AHG is added and then centrifuged. Anti-Human Globulin AntiIgG, -C3d; polyspecific acts as a link between the antibody and/or complement coating of neighbouring red blood cells and induces agglutination. Uncoated red blood cells will not agglutinate. No agglutination or hemolysis indicates compatibility Detects Anti-Fya, -Jka, -K Antibodies present in acquired hemolytic anemia Antibodies in Rh system which react only in the AHG test (called 3rd order or cryoagglutinoid antibodies) Incorrect ABO grouping of the patient or donor An alloantibody in the patients serum reacting with the corresponding antigen on donors RBCs An autoantibody in the patients serum reacting with the corresponding antigen on donors RBCs Bacteriogenic pan agglutination (Huebener-Thomsen Friedenreich phenomenon). Reaction takes place at 20C not at 37C. Non-bacteriogenic pan agglutination caused by acquired hemolytic anemia or by rare specific antibodies. Reaction takes place at 37C. DAT is always (+). Prior coating of the donors RBCs with protein, resulting in positive AHG test Abnormalities in the patients serum Albumin/Globulin (A/G) ratio imbalance Presence of high molecular weight dextrans or other plasma expanders Antibody against additives in the albumin reagents

Problems Encountered
Rouleaux formation In some diseases myclomatosis macroglobulinemia Certain synthetic plasma expanders like dextran Fibrinogen

Pan agglutination spontaneous clumping of cells against a given serum

Causes of Positive Results in Crossmatching

Polyagglutinability Cold agglutinins Whartons jelly Prozones Ag-ab deterioration Presence of other immediate spin-reactive antibodies

CROSSMATCHING Albano, Astorga, Dumangon, Fregil, Gutierrez, Loque 4AMT

Hyperimmune ABO antibodies Procedure is not performed properly Infants specimens are tested Contaminants in the test system SOLUTION: EDTA has been reported to eliminate some of the false-positive reactions

Procedures to shorten cross-matching time

Test should be performed at 37C LISS instead of saline 2% suspension of donor red cell Enhancing agents (albumin, enzymes) Polycation or polybrine

CASE

PROCEDURAL PHASE Protein Phase Thermo Phase AHG Phase Protein Phase Thermo Phase AHG Phase Protein Phase Thermo Phase AHG Phase Protein Phase Thermo Phase AHG Phase

CROSSMATCH MAJOR MINOR 0 0 0 0 0 0 + 0 0 0 + +

INTREPRETATION Blood is compatible in both major and minor crossmatch. Blood is safe for transfusion. Blood is incompatible in major crossmatch but compatible in minor crossmatch. Blood is not safe for transfusion Blood is incompatible both in major and minor crossmatch. Blood is not safe for transfusion.

Blood is compatible in major crossmatch but incompatible in minor crossmatch. Blood can be 0 transfused but with caution. Table 1.1 Summary of Interpretation of Crossmatching Results

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