Crossmatching - it is the testing of patient's blood against donor's blood - only part of pretransfusion compatibility testing Two Main

Functions: 1. 2. Final check of ABO compatibility between donor and patient May detect the presence of an antibody in the patients serum

Procedure in Crossmatching a. Protein/Albumin/Room Temperature Phase (Immediate Saline Spin) IgM related incompatibilities Thermo/Incubation Phase (37 C) IgG related incompatibilities Antihuman Globulin Phase/Coombs Phase (AHG) Unexpected antibodies that are complement activated

b. c.

Kinds of crossmatch: a) Major crossmatch (PS-DR) - a type of crossmatch that involves mixing of patient's serum and donor's red blood cells. -this is much more critical for ensuring safe transfusion than minor test. b) Minor crossmatch (PR-DS) - a type is crossmatch that involves mixing of patient's red blood cells and donor's serum. c) Immediate Spin Saline crossmatch (Simplest Serologic Crossmatch) - a process of mixing the recipient's serum with donor's red blood cells and centrifuging immediately. Absence of hemolysis and agglutination indicates compatibility. - designed to detect ABO incompatibilities d) Computerized crossmatch - crossmatch performed by a computer. - designed to detect ABO incompatibility. -compares recent ABO serologic test results and interpretation on file for both donor and patient being matched and determines compatibility based on comparison. e) Abbreviated crossmatch - a type and screen coupled with immediate spin. -Type and screen is a policy in which the patient's blood sample is tested for ABO, Rh, and unexpected antibodies, then stored in the blood bank for immediate crossmatching. 2 f) Antiglobulin Crossmatch - procedure begins in the same manner as the immediate spin crossmatch but it continues to 37C incubation and finishes with an Antiglobulin test. g) Autocontrol - consists of patient's own cells and serum and may be tested in parallel with the crossmatch (PS-PR) Protein Phase Thermo Phase AHG Phase + O O O Interpretation of Crossmatching Results: Compatible = if there is no agglutination or hemolysis in all tubes Incompatible = if there is presence of agglutination or hemolysis in any tubes Case 1 Procedural Phase Protein Phase Thermo Phase AHG Phase Crossmatch Major Minor O O O O O O Interpretation Blood is compatible both in 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

Protein Phase Thermo Phase AHG Phase

Protein Phase Thermo Phase AHG Phase

O O O

Blood is incompatible in both major and minor crossmatch. Blood is not safe for transfusion Blood is compatible in major crossmatch but incompatible in minor crossmatch. Blood can be transfused but with caution

If one isolated positive result is obtained then a DAT should be performed on the donors RBCs Donor cells that demonstrate a (+) DAT will be incompatible with all recipients tested in the AHG phase o Because the cells are already coated with immunoglobulin and or complement

5.

Abnormalities in the patients serum A. Imbalance of the normal ratio of albumin and gamma globulin (A/G ratio) Diseases as multiple myeloma and macroglobulinemia May cause RBCs to stick together on their flat sides o Appearance: stacks of coins o Rouleaux formation Strong rouleaux may mimic true agglutination Refractile Strongest after 37 C incubation test Problems with rouleaux may be resolved using saline replacement technique Presence of high molecular weight dextrans or other plasma expanders May cause false positive results in compatability Saline replacement may resolve the problem

Causes of Positive Results in the Serologic Crossmatch 1. Incorrect ABO grouping of the patient or donor ABO grouping should be repeated An alloantibody in the patients serum reacting with the corresponding antigen on donor RBCs Autocontrol tube should be negative o Unless the patient has been recently transfused with incompatible RBCs Antibody screening test (+) o Identification of antibody specificity a. If RBCs of all donors tested are incompatible with the patients serum and the antibody screening test is (+) : = antibody directed against an antigen of high incidence or multiple antibodies in the patients serum If antibody screening test (-) and only one donor unit is incompatible = antibody in the patients serum may be directed against an antigen of relatively low incidence that is present on that donors RBCs 6. c. If antibody screening test (-) = Patients serum may contain naturally occurring or passively acquired ABO agglutinins

2.

B. -

C. -

b.

An antibody against additives in the albumin reagents May cause false-positive results in compatibility testing Rarely a patients serum reacts against the albumin in testing reagents o Occurs when the patient has antibodies stabilizing substances such as caprylate added to the albumin reagents

Contaminants in the test system Dirty glassware Bacterial contamination Chemical or other contaminants in saline Fibrin clots

3.

An autoantibody in the patients serum reacting with the corresponding antigen on donor RBCs Auto-control tube will be (+) Antibody screening test and tests of the patients serum with donor cells: (+) Prior coating of the donor RBCs with protein, resulting in a positive antihuman globulin test

= may cause false-positive results

4.

Observations Crossmatch: (+) Auto-control: (-) Antibody Screen: (-)

Crossmatch: (+) Auto-control: (-) Antibody Screen: (+)

Crossmatch: (+) Auto-control: (+) Antibody Screen: (-)

Investigation of Incompatible Major Crossmatches Possible Interpretations Comments Incorrect ABO grouping of patient or Repeat ABO grouping: verify identity of the donor sample Patients serum may contain ABO Check patients sample for subgroups; check antibody patients transfusion and transplantation histories Alloantibody in patients serum Perform antibody identification tests on patients reacting with antigen donors red serum and repeat crossmatch using units cells but not present on screening negative for the corresponding antigen. If studies cells are noninformative and patient I incompatible with only 1 unit, locate other compatible units Donor unit may have a positive DAT Perform DAT on donor unit; if positive, do not use the unit. Alloantibody in patients serum reacting with antigens on donors Perform antibody identification studies on cells and screening cells patients serum and repeat crossmatch using units negative for the corresponding antigen If unable to identify antibody specificity, consult a reference laboratory. Both an autoantibody and Perform autoadsorption of patient serum to alloantibody may be present in the remove autoantibody (if not recently transfused), patients serum perform antibody identification tests, repeat compatibility tesets using autoadsorbed serum Abnormalities in the patients serum owing to imbalance of A/G ratio: If rouleaux are seen, use saline replacement Plasma expanders technique Caprylate antibodies Obtain new specimen Contaminants Use caprylate-free reagents Repeat tests using fresh saline, new bottles of reagents, clean test tubes.

Sources: Harmening, D. M. (2005). Modern blood banking and transfusion practices. (5th ed., pp. 271-272). Philadelphia, Pennsylvania: F.A Davis Company. Cardona, C. C., Martin I, G. L., & Garcia, R. S. (2011).Laboratory manual in blood banking. (pp. 46-48). Manila, Philippines: C&E Publishing, Inc.