Guide Questions:

1.

What is detected by the immediate spin phase?

Designed to detect compatibility of IgM type of antibodies in patient's serum against

antigen on donor red cells in saline phase, i.e. ABO compatibility testing.
IgM antibodies react best at cold temperatures.
An immediate spin, saline major crossmatch used to determine ABO compatibility in
patients with no demonstrable clinically significant antibodies and a negative history of
antibody formation. Under some conditions, this determination of ABO compatibility can
also be accomplished by an electronic crossmatch.
This is performed if:
a. There is no clinically significant unexpected antibodies detected in the antibody screen
using reagent red blood cells that are not pooled.
b. There is no record of previous detection of clinically significant antibodies.
2.

What is detected by the warm saline phase?

Where the immunologic reaction between red cells suspended in saline medium and the
antibody occurs at room temperature.
IgG antibodies react best at warm temperatures, and no visible agglutination commonly
seen.
3.

What is detected by the high protein phase

Where the red cells are suspended in the antibody (serum) with 22% albumin (protein)
and incubated for 30 min at 37C. The high protein environment enhances agglutination of
univalent antibodies such as Rh group. The modern use of LISS has helped to reduce the
incubation time for 30 to 15 min. use of LISS eliminates the use of albumin.
The high protein phase can demonstrate antibodies in the Rh system. Presence of
agglutination or hemolysis indicates incompatibility. The high protein phase does not only
strengthen or enhance the reactions but also reveals antibodies not demonstrable by the
saline-antiglobulin test.
Causes agglutination by adjusting zeta potential between RBCs. Detects IgG type of
antibody.

4.
What is detected by the AHG phase?
In addition to saline phase compatibility testing (IgM antibodies detection), AHG testing is
designed to detect compatibility of IgG type of antibodies in patient's serum against antigen
on donor's red cells.
AHG phase involves the addition of antiglobulin sera to the incubated test tubes. With this
addition, antihuman antibodies present in the sera become attached to the antibody
globulin in the RBCs, causing agglutination. The antiglobulin phase detects most incomplete
antibodies in the blood group systems, including Rh, Kell, Kidd and Duffy BGS.
AHG acts as a bridge cross-linking red cells sensitized with IgG antibody or complement.
AHG has specificity for the FC portion of the heavy chain of the human IgG molecule or
complement components.
Modern Blood Banking and Transfusion Practices
By Denise M Harmening
Page67-69

Post Lab Conference Questions:

1.
How is an incompatible cross match resolved or further investigated?
a. ABO grouping should be immediately repeated, especially if strong compatibility is
observed in a reading taken after immediate spin.
b. If the antibody screening test is positive, antibody identification panel studies would
allow antibody specificity; which then permits selection of units lacking the antigens
for compatibility testing
c. Monitor the patient for signs of delayed hemolysis
d. A DAT should be performed on the Donors RBC
e. Problems with rouleaux can be often resolved using the saline replacement
technique.
f. Compatibility testing could then be performed using the autoabsorbed serum.
(autoadsorption of the patients serum to remove autoantibody activity)
2. What are the different precautions and sources of error in the serologic cross match?
Precautions:

Handle the sample gently to prevent hemolysis, which can mask hemolysis, which
can mask hemolysis of the donors RBCs.

Label the sample with the patients name, the hospital or blood bank number, the
date, and the phlebotomist initials.


Indicate on the laboratory request the amount and type of blood component needed.

If more than 72 hours have elapsed since an earlier transfusion. Previously

crossmatched donor blood must be crossmatched with a new recipient serum sample to
detect newly acquired incompatibilities before transfusion.

If the patients scheduled for surgery and has received blood during the past 3
months, be aware that his blood needs to be crossmatched again if his surgery is
rescheduled to detect recently acquired incompatibities.
Sources of error:
1.
Incorrect ABO grouping of the patient or donor.
2.
All alloantibody in the patients serum reacting with the corresponding antigen
on donor RBCs.
3.
An auto antibody in the patients serum reacting with the corresponding antigen
on donor RBCs.
4.
Prior coating of the donor RBCs with protein, resulting in a positive anti human
globulin test.
5.
Abnormalities in the patients serum.
6.
Contaminants in the test system
3.
What are the different types of cross matching?
There are two types of cross matching (i) major cross matching and (ii) minor cross
matching.
Major cross matching, the cells of the donor are directly matched against the plasma of
recipient. It is important to ensure that antibodies present in the recipients plasma do not
harm the donors red cells.
Minor cross matching, the donors plasma is checked against the red cells of the recipient. It
is called minor cross matching against because it is not so important, as the small volume of
the donors plasma is diluted in a large volume of the recipients plasma. Therefore, the
titre of antibodies present in the donors plasma falls to such a low level after transfusion
that they are quite unlikely to damage the red cells of the recipient.
Reference:
Nursing: Deciphering diagnostic tests: edited by Lippincott Williams & Wilkins page 177;
Modern Blood Banking and Transfusion Practices
By Denise M Harmening
Page 249
Textbook Of Practical Physiology - 2Nd Edn.
By G.K. & Pal, Pal, Pravatipg 98

Angeles University Foundation

College of Allied Medical Professions

Written Report in
Blood Banking (Lab)
Crossmatching

Submitted By:
Bautista, Jericho
Escoto, Jannie Hazel
Pineda, Katreena Marie
Serrano, Sheena
Submitted to:
Mrs. Engracia Arceo
Mr. Michael Tanglao