A C T I V I T Y 4 B
REVERSE ABO
BLOOD TYPING TUBE METHOD
PRESENTED BY GROUP 4,
BLOCK BB; A.Y. 2023-2024
introduction
Reverse typing identifies the antibodies present in a patient's
serum or plasma by reacting the patient's serum with red cell
suspension of known groups A or B cells and then observing for
agglutination. The principle follows that whenever a blood group
antigen is present on the red cell, the opposite antibody is present
on the serum. Reverse grouping should be performed using a test
tube. This is a sensitive test tube method recommended to enhance
agglutination with weakly reacting serum specimen.
objectives I DETERMINE THE ANTIBODIES PRESENT FROM
THE SERUM SAMPLE OF AN INDIVIDUAL BY
USING KNOWN RED CELLS;
II DETERMINE THE IMPORTANCE OF
PERFORMING REVERSE ABO
TYPING; AND
IV IDENTIFY THE COMMON SOURCES OF
TECHNICAL ERRORS THAT MAY RESULT
IN ABO DISCREPANCIES.
MATERIALS
Test tubes
Test tube rack
Serum sample to be tested
5% red cell suspension of Known group A cells
5% red cell suspension of Known group B cells
Pasteur pipettes
NSS
Clinical centrifuge
PROCEDURE
1. Prepare 5% suspension of A 2. Label 2 tubes "A" and "B".
and B red cells.
PROCEDURE
3. Place 2 drops of serum 4. Add 1 drop of red cell suspension
sample to each tube. A to "A" tube and 1 drop of B cell
suspension to"B" tube.
PROCEDURE
5. Mix gently and centrifuge 6. Observe the supernate for
for approximately 1 minute. hemolysis.
PROCEDURE
7. Gently dislodge the cell button and examine
for the presence of agglutination with the aid of
a viewing box or place a drop of the solution
on a slide and read under the low power
objective lens of the microscope.
NOTE: Weak reactions will be enhanced if the
tubes are left for more than 5 minutes at room
temperature before centrifugation. Avoid
excessive shaking which may disperse weak
agglutination.
Hemolysis indicates the presence of antibodies
thus mean a positive test.
RESULTS
A
Mix gently and centrifuge for Observe the supernate for
approximately 1 minute. hemolysis.
B
Mix gently and centrifuge for Observe the supernate for
approximately 1 minute. hemolysis.
O
Mix gently and centrifuge for Observe the supernate for
approximately 1 minute. hemolysis.
AB
Mix gently and centrifuge for Observe the supernate for
approximately 1 minute. hemolysis.
REVIEW
QUESTIONS
When When there is hemolysis in the final solution it
indicates that an incompatibility has occurred
hemolysis between the patient's serum and the known
is present
red blood cells (RBCs) used in the test. When
incompatible antibodies in the patient's serum
in the final
encounter the corresponding antigen on the
known RBCs, they bind to them, triggering the
solution, destruction of the RBCs. This release of
hemoglobin from the lysed RBCs causes the
what does solution to appear red and clear (hemolyzed).
This hemolytic reaction signifies the presence
it indicate? of antibodies in the patient's serum targeting
Explain.
the specific antigen present on the known
RBCs used in the test.
This can happen in:
Newborns: Their immune systems are still
When do
developing, and they haven't been exposed
to enough antigens to produce strong
ABO antibodies.
Immunosuppressed individuals: Medications
discrepancie or medical conditions can weaken the
s occur? immune system's ability to produce
antibodies.
How can Congenital immunodeficiency: Individuals
may have a genetic condition affecting their
these be antibody production.
resolved? Resolved by:
Investigating the cause
Repeating the Test
This can happen in:
When do
Newborns: Their immune systems are still
developing, and they haven't been exposed
ABO to enough antigens to produce strong
antibodies.
discrepancie Immunosuppressed individuals:
s occur?
Medications or medical conditions can
weaken the immune system's ability to
How can produce antibodies.
Congenital immunodeficiency: Individuals
these be may have a genetic condition affecting
resolved? their antibody production.
Technical errors: Mistakes during blood
collection, sample handling, or test
procedures can lead to inaccurate result.
Sample collection from the wrong patient
What are the Labeling errors
common Failure to add reagents
sources of Failure to add sample
technical Using expired or contaminated reagents
Adding reagents or sample in wrong
errors that quantities
may result in Transcription errors
ABO Using the wrong sample
discrepancies Uncalibrated centrifuge
? Warming during centrifugation process
Mis-reading of strength of reaction
thank
you
References:
LearnHaem. (2020, April 6). ABO Discrepancies. LearnHaem | Haematology Made Simple.
https://www.learnhaem.com/courses/frcpath-transfusion/lessons/blood-grouping-
2/topic/abo-discrepancies/
