ANTIBODY SCREENING AND IDENTIFICATION • The fluid/supernatant to which the red blood cells are suspended can’t pass

By: Donald Gerniel Eborde, RMT through the gel

OBJECTIVES: o Solid Phase Adherence Method
1. Be able to know the difference between antibody screening test and antibody identification ➢ RBC Reagents
test o Group “O” cells, typed with the most common and most significant antigens
2. Be able to know the significance of performing these tests o A.k.a screen cells
3. Be able to know how to isolate antibodies if present o Packaged in 2 or 3 cell suspensions each having a unique combination of antigens
4. Be able to perform the tests satisfactorily o D, C, c, E, e, K, k, Fya, Fyb, Jka, Jkb, Lea, Leb, P1, M, N, S, s
PRINCIPLE: o Each screen cells will be accompanied by antigen profile sheet, detailing which
These tests detect antibodies in serum which react against red cell antigens. If the antibody antigens are present in each vial of cells
screening test yields a positive result, this means that antibody/ies is/are present in the serum o Homozygous expression of antigen is ideal
and needs to be identifies or isolated, thus one must proceed to antibody identification test. ▪ More antigens will be expressed on red blood cells
➢ Makes use of Indireact Antiglobulin Test (IAT) ▪ Weak reacting antibodies will be detected
➢ What type of crossmatching was omitted because of antibody screening? Minor o Jk, Fy, M, N
crossmatching ▪ Antigens on screen cells that actually show dosage
➢ Antibody screening is part of the routine testing • “DOSAGE” – difference of expression of antigen between homozygous
➢ The detection of antibodies directed against red blood cell antigens is critical in and heterozygous
pretransfusion compatibility testing ➢ “Autocontrol”
➢ Focuses on “irregular” or unexpected antibodies o Patient serum vs patient red cells
➢ UNEXPECTED ANITBODIES o Normally, should be negative
o Immune alloantibodies ▪ If positive, usually related to DAT positive
▪ Counterpart of naturally occurring antibody; antibodies produced through ▪ Some medications also cause positive DAT/autocontrol
stimulation
o Naturally occurring antibodies
▪ Produced without stimulation
• i.e ABO (anti-A, anti-B)
o Passively acquired antibodies
▪ Vaccines, intravenous immunoglobulin, Rhogans (produced for Rh negative
mothers to prevent HDN) ANTIBODY SCREEN VS CROSSMATCH
➢ CLINICALLY SIGNIFICANT ANTIBODIES:
o IgG
Antibody Screen Crossmatch
o 37°C or AHG -Screen cells set will have cells with - Donor units may or may not have
➢ Required to detect clinically significant antibodies in both the blood donor and the homozygous expression, making it homozygous antigen expression,
intended recipient as part of pretransfusion compatibility testing more reliable in detecting weakly possibly not detecting weakly reacting
➢ Only a small percentage of the population (0.2%-2%) has detectable RBC antibodies reacting antibodies antigens
➢ METHODS: - More superior than crossmatching
o Tube Method
▪ Saline phase/Immediate phase
• Now an option because in this phase, you will detect clinically
insignificant antibodies which don’t react at 37 degree Celsius
▪ Protein Phase
▪ AHG phase ANTIBODY SCREEN INTERPRETATION
o Gel Method
▪ Doesn’t require washing of red blood cells
 ➢ Patient’s serum or plasma is tested against additional RBCs possessing known antigens

ANTIBODY IDENTIFICATION PANEL

➢ A collection of 11-20 group “O” RBCs with various antigen expression
o “O”RBCs are used to prevent interferences against anti-A and anti-B
➢ The method used in antibody screening should be the same method to be used in
antibody identification – for standardization
➢ Evaluation of Panel Results
o Exclusion a.k.a “Rule-Out” ➢ With the exception of ABO antibodies, the presence of antibodies either in the serum or
1. Check the negative reaction (a + check cell) on the surface of the red cells is an unexpected finding in the blood bank. It is, however, a
• Negative autocontrol – no autoantibodies present situation that may influence the successful outcome of a transfusion. Along with ABO and
• + signs – indicated that the antigen is present Rh testing, a test is performed in the blood bank to detect antibodies in the serum is the
2. Cancel antigen that has “+” antibody screen test. For a more definitive test and for the isolation of the antibody
• These antigens are not the cause of the positive reaction identification test or panel is performed.
3. Check which reaction phase is positive
• If only one phase is positive, most probably, only one antibody is causing
the reaction
• If more than 2 or 3 phases are positive, 2 or more antibodies are present
[Link] the strength of the reaction
• Most probably, the antibody showing dosage
• If same reaction – 1 antibody
[Link] the positive reaction
• Cancel antigens with zero (“0”)
[Link] not cancelled is theunknown antbody
o Inclusion
▪ Autocontrol ➢ A second test is done to detect antibodies and that is the DAT. It detects antibodies
▪ Phase of reaction and strength coating the surface of the red cells in vivo. These antibodies are also identified by using an
antibody identification or panel but can only be accomplished after the antibodies have
been removed from the cells by a process called ELUTION.
➢ As technology advances, one of the most commonly used antibody identification these
days employ the gel technology. It has a gel incorporated in its system with pores in it.
When there is agglutination, the clumping cannot pass through the pores of the gel in the
system thus it will stay on the top after centrifugation. On the other hand, when there is no
agglutination, after centrifugation, the cells can pass through the gel pores and therefore
settle at the bottom of the system.