Compatibility Testing

❖ The transfused red cells will have an acceptable survival

Outline
rate, and there will be no significant destruction of the
• Collection and Preparation Sample recipient’s own red cells.
• Collecting Patient Samples
• Donor Samples Collection and Preparation of Samples.
• Selection of Appropriate Donor Units
• Testing of the donor sample • Positive Patient Identification
• Testing of the patient sample The major cause of transfusion associated facilities
• Cross match have been clerical errors resulting in incorrect ABO
• Summary of Pre-transfusion Testing grouping.

The most common cause of clerical errors and thus

transfusion accidents is misidentification of the
❖ As the knowledge of the new blood group system increased,
patient involved in the transfusion. Confusion in
so did the search for more sensitive pre-transfusion
identification of the patient when the blood sample
compatibility testing methods. Pioneer blood bankers mixed
was drawn a mixed up samples during handling in
the patient’s serum and the donor’s red cells and observed
the lab, and error in identification of he patient
for direct red blood cell lysis, agglutination, or both. This
when the transfusion was given.
became known as the major cross-match test.

❖ The term compatibility test and cross-match are sometimes ❖ To prevent collection of the samples from the wrong patient
the blood request form must be used to confirm patient’s
used interchangeably, they should be clearly differentiated.
identity before phlebotomy is performed. The request form
must state the intended recipient’s full name, and unique
❖ The cross-match became part of a series of pre-transfusion
test known as compatibility testing. The compatibility test hospital identification number.
includes an ABO and Rh grouping performed on the donor
❖ Other information such as age and date of birth, address,
and recipient samples, screening antibodies and a cross-
sex, and name of requesting physician can be used to verify
match.
patient identity further bit is not required on the form.
❖ The purpose of pre-transfusion or compatibility testing is to
❖ The patient’s wrist band identification must always be
ensure the best possible results of a blood transfusion.
compared with the requisition form.
 the phlebotomy, and the numbers must be verified again
❖ If the patient does not have a wrist band or if the patient’s immediately after filling.
identity is unknown, some form of positive identification
must be attached to the patient before collection of ❖ Donor and recipient samples must be stored for a minimum
samples. of 7 days following transfusion.

Collecting Patient Samples

❖ The samples should be stoppered and refrigerated at 1- 6°C,
Hemolyzed samples can not be used for testing because carefully labeled, and adequate in volume so that they can
hemolysis caused by activation of complement. be re-evaluated if the patient experiences an adverse
response to the transfusion.
Serum or plasma may be used for pre-transfusion testing.
Most blood bank technologist prefer serum because plasma
Selection of Appropriate Donor Units.
may caused small fibrin clots which may difficult to
distinguish from true agglutination. In almost all cases, blood and blood components of the
patient's own ABO and Rh group should be selected for
❖ When specimen is received in the lab, a blood bank transfusion.
technologist must confirm that the information on the
When blood and blood components of the patient's type
sample and requisition form agree.
are unavailable or when some other reason precludes their
use, units selected must lack any Ag against which the
❖ All discrepancies must be resolved before the sample is
patient has a significant Ab.
accepted, and if any doubt exists, a new sample must be
drawn. ❖ When transfusion of an ABO group different from the
recipient must be given, packed red cells must be used
Donor Samples.
rather than whole blood which contains plasma Abs that are
Donor testing samples must be taken when the full donor incompatible with the patient's red blood cells.
unit is drawn. Depending on the method used for testing, ❖ Group O packed red blood cells can be safely used for all
clotted sample, anti-coagulated samples, or both, are patients, however, conservation of a limited supply of group
obtained. O blood should dictate its use for patients of other AB types
only in special circumstances.
Donor information and medical history card, the pilot
samples for processing, and the collection bag must all be
labeled with the same unique number code before starting
 ❖ If ABO-specific blood is not available or is in less than prevent disease transmission must be performed on a
adequate supply, alternative blood groups are chosen as sample of blood taken at the time of collection of the unit of
summarized in the following table; blood from the donor.

Patient’s BG Alternative BG given as packed cells ❖ A screening test for unexpected antibodies to red blood cell
O NONE Ags is required by AABB standards on samples from donors
A O revealing a history of prior transfusion or pregnancy.
B O
Testing of the donor sample
AB A, B, O
❖ The transfusing facility is required by AABB standards to
Only one of the three should be used for a given patient
confirm the ABO cell grouping on all units and Rh grouping
❖ Rh-negative blood can be given to Rh-positive patients, on units labeled Rh-neg.
however, good inventory management again should
conserve this limited resource for use in Rh-neg recipients. ❖ Tests for weak D (Dч) are not required to be reported. The
transfusion facility does not need to repeat any other
❖ If Rh-neg units is near expiration, the unit should be given testing procedure.
rather than wasted.
Testing of the patient sample
❖ Rh-pos blood should not be given to Rh-neg women of ❖ A record of all results obtained in testing patient samples
childbearing age. must be maintained.
❖ Identification number should be assigned each time a
❖ Transfusion of Rh-neg male patients and female patients patient is admitted for treatment.
beyond menopause with Rh-pos blood is acceptable as long ❖ Any discrepancies between previous and current results
as no performed anti-D is demonstrable in the sera. must be resolved before transfusion is initiated.
❖ A new sample should be collected from the patient, if
necessary to resolve the problem.
Compatibility Testing Protocols
❖ ABO and Rh grouping should be included in the file.
Testing of the donor sample. ❖ Also, notations concerning unusual serologic reactions and
the identity of unexpected Abs in the patient’s serum should
❖ According to the Code of Federal Regulation (CFR) and the
be included.
American Association of BB (AABB) standards, ABO and Rh
grouping (including a test for weak D) and tests intended to
❖ ABO and Rh grouping and Ab screening of the patient serum can autoantibody, (alloantibodies, if the patient’s has been recently
be performed in advance of or at the same time as the cross- transfused) is responsible for the positive control result.
match.
❖ If the Rh group of the recipient can not be determined and
❖ If the patient has had a transfusion or has been pregnant within transfusion is essential, Rh negative blood should be given.
the last 3 months or if the history is unavailable or uncertain,
the sample must be obtained from the patient within 3 days of ❖ The test for Dч is unnecessary when testing transfusion
scheduled transfusion. recipients. Individuals typing as Rh neg in direct testing should
receive Rh-neg blood and those typing as Rh pos in direct
ABO Grouping testing should receive Rh pos blood.
❖ Determination of the patient’s correct ABO group is the most
critical pre-transfusion serologic test. ❖ As Dч are considered Rh pos and may receive Rh pos blood
during transfusion.
❖ If the cell and serum grouping do not agree, additional testing
Antibody Screening
must be conducted to resolve the discrepancy.
❖ The patient’s serum or plasma must be tested for unexpected
❖ If the patient’s ABO group cannot be satisfactory determined Abs.
and immediate transfusion is essential, group O packed red
blood cells should be used. ❖ The aim of the Ab screening test is to detect as many clinically
significant Abs as possible.
Rh Grouping
❖ Rh grouping is performed using anti-D blood grouping serum. ❖ Clinically significant Abs refers to Abs that are reactive at 37°c or
Tube or slide test should be performed according to the in the DAT or both and are known to have caused a transfusion
manufacture’s directions for the reagent, which may or may not reaction or unacceptably short survival of the transfused red
include the use of a suitable diluents control. blood cells.

❖ Control must be run in parallel with Rh grouping test performed Testing of the patient sample
on patient’s samples, to avoid incorrect designation of Rh neg, ❖ Abs Regarded as always being potentially clinically
patient as Rh positive. significant
ABO Rh Kell Duffy Kidd S s U
❖ Direct antiglobulin test (DAT) should be performed on the
patient’s red blood cells to determine whether uptake of ❖ Abs that may sometime be clinically significant
 Lea p Lua Lub Cartwright ❖ 2 – Testing can be performed well in advance of the
anticipated transfusion, allowing ample time for
❖ Abs that rarely, if ever, are clinically significant identification of unexpected Ab and location of
Leb Chido/Rodgers (Cha / Rha ) York, Sd Xg& Bg suitable donor units lacking the corresponding Ag
❖ Methods used to detect Abs in patient’s sera must
❖ Correct ABO grouping results are much more critical to demonstrate all significant coating, hemolyzing, and
transfusion safety than Ab screening. agglutinating Abs active at 37°c.

Cross-match
❖ Most Abs, other than anti-A and anti-B do not cause severe
hemolytic transfusion reactions. Thus the vast majority of ❖ Two main functions of the crossmatch test can be cited as,
patients would not suffer grave consequences if transfused with
blood from ABO group compatible donor without the benefit of i. It is the final check of ABO compatibility between
Ab screening tests. donor and patient.
❖ Detection of unexpected Abs is important, however, for the
selection of donor red blood cells that are likely to survive ii. IT may detect the presence of an Ab in the patient’s
maximally in the patient circulation. serum that will react with Ags on the donor RBCs
but that was not detectable in the Ab screening
❖ Weakly reactive Abs that are capable of reacting with their because the corresponding Ag was lacking from the
Ags at 37°c can cause decreased survival of transfused screening cell.
incompatible red cells.

❖ Because large numbers of Ab molecules are present in the

patient’s circulation compared with the number of red cells Major and Minor cross-match test
in a unit of blood, incompatible donor cells are highly ❖ Major cross-match test, consisting of mixing the patient’s
vulnerable to destruction by patients Abs. serum with donor RBC’s.

❖ Abs screening offers several advantages over direct cross- ❖ Minor cross-match test, consisting of mixing the donor’s
matched testing for detection of Abs; plasma with patients RBCs.
❖ 1 – Testing is performed using selected group O red
cells that are known to carry optimal representation ❖ The minor cross-match test has been completely eliminated
of important blood group Ags. in most blood banks, because donor’s samples are screened
beforehand for the more common Abs.
 ❖ The procedure begin in the same manner as the IS cross-
match, continues to 37°c incubation and finishes with AHG
Method for major cross-match test.
test.
❖ Cross-match methods can be categorized by the test phase
in which the procedure ends.

❖ Immediate spin (IS) cross-match (Abbreviated Cross-match)

❖ When no clinically significant Abs are detected nor are there

previous record of such Abs, a serologic test to detect ABO
incompatibility is sufficient.

❖ False reacting may be seen in the presence of other IS

reaction (auto-I). In patient with hyperimmune ABO Abs,
when the procedure is not performed correctly (delayed in
centrifugation or reading) when rouleaux is observed, or
when infant’s specimens are tested. Pre-transfusion Testing

❖ It is to provide safe, compatible blood for transfusion to

Antiglobulin Cross-match each individual patient. The necessary for safe transfusion
are:

❖ 1. Accurate ABO and Rh typing of the patient.

❖ 2. Accurate ABO and RH typing of the donor.
❖ 3. Screening tests for antibodies in the donors and patient’s
serum.
❖ 4. In the presence of patient’s antibodies, selection of
appropriate units for each patient.
❖ 5. Compatibility Testing – (Major)
❖ 6. Accurate completion of paperwork and labels

Compatibility Testing
 Each compatibility test is a unique experiment in which an ❖ Prove that donor and or recipient serum is free of
unknown (patient) serum and (donor) red cells are tested for the antibodies. Prevent immunization of the recipient.
detection of unexpected antibodies which are directed against ❖ Detect ALL ABO typing errors.
antigens found on the cells. Negative results indicate compatibility. ❖ Detect errors in Rh typing of either recipient or donor unless
This is one of the most important tests performed by a transfusion the recipient’s serum contains an Rh antibody.
service. ❖ Detect ALL error of identification.
❖ Pre-transfusion testing of the recipient must include an ABO
The purposes of compatibility testing are:
and Rh typing, antibody screen. And a cross-match with all
❖ 1. To detect irregular antibodies in the recipient serum that donor units.
are directed against the donor’s cells.
❖ 2. To detect errors in ABO grouping.
❖ 3. To detect errors in labeling, recording, or identifying
donor’s or reporting samples.

Compatibility testing does not:

❖ 1. Ensure normal survival of donor red cells.