Serology of Blood and Blood Typing

Clinical Biochemistry
M.Sc-Biochemistry
17-10-2018

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History: Karl Landsteiner
• Discovered the ABO
Blood Group System in
1901
• Blood types:
Type A, Type B, Type AB,
& Type O

The identification of blood

group depends on the
antigen and antibodies
produced by the immune http://www.nobelpreis.org/castellano/medizin/images/landsteiner.jpg
system.
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 Blood Grouping
• Transfusion is the transfer of blood or blood components from one individual to another.

• Infusion is the introduction of fluid other than blood, for example: saline solution or glucose

solution.

• Blood Group is determined by the antigens on the surface of RBCs.

• Antibodies can bind to RBC antigens, resulting in agglutination (clumping) or hemolysis (rupture) of

RBCs

• Blood Groups: ABO

• Rhesus (Rh) factor is an inherited protein found on the surface of red blood cells. If your blood has

the protein, you're Rh positive. If your blood lacks the protein, you're Rh negative. Rh positive is the

most common blood type.

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ABO Blood Typing
Blood Antigens (Agglutinogens) on Antibodies
Type Red Blood Cells (Agglutinins)
in Plasma

A A Anti-B

B B Anti-A

AB A&B None

O Neither Anti-A & Anti-B

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ABO Basics
• Blood group antigens are actually sugars attached to the red
blood cell.
• Individuals inherit a gene which codes for specific sugar(s) to
be added to the red cell.
• The type of sugar added determines the blood group.

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RBC Precursor Structure

RBC

Glucose

Galactose
Precursor
Substance
(stays the N-acetylglucosamine
same)
Galactose
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Formation of the H antigen
The H gene codes for
an enzyme that adds
the sugar fucose to the RBC
terminal sugar of a
precursor substance

Glucose

H antigen Galactose

N-acetylglucosamine

Galactose
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Fucose
Formation of the A antigen
The “A” gene codes for an
enzyme (transferase) that adds RBC
N-acetylgalactosamine to the
terminal sugar of the H antigen
N-acetylgalactosaminyltransferase
Glucose

Galactose

N-acetylglucosamine

Galactose
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N-acetylgalactosamine
Fucose
Formation of the B antigen

The “B” gene codes for RBC

an enzyme that adds D-

galactose to the terminal
sugar of the H antigen Glucose

D-galactosyltransferase
Galactose

N-acetylglucosamine

Galactose
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Galactose
Fucose
Group O
• Approximately 45% of the
population is group O.
• No A or B antigens present, think
of as “0” antigens present.
• These individuals form potent anti-
A and anti-B antibodies which
circulate in the blood plasma at all
times.
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Group A
• Approximately 40% of the
population is group A.
• No B antigens present.
• These individuals form
potent anti-B antibodies
which circulate in the blood
plasma at all times.

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Group B
• Approximately 11% of the
population is group B.
• No A antigens present.
• These individuals form potent
anti-A antibodies which circulate
in the blood plasma at all times.

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Group AB
• Approximately 4% of the
population is group AB.
• Both A and B antigens
present.
• These individuals possess no
ABO antibodies.

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Hemolysis
• If an individual is transfused with an incompatible
blood group destruction of the red blood cells will
occur.
• This may result in the death of the recipient.

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Summary

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 Summary

Blood Group Antigens on cell Antibodies in Transfuse with

plasma group

A A Anti-B A or O

B B Anti-A B or O

AB A and B none AB, A, B or O

O None Anti-A & B O

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Rh Factor
Rhesus (Rh) factor is an inherited protein found on the surface of

red blood cells. If your blood has the protein, you're Rh positive. If

your blood lacks the protein, you're Rh negative. Rh positive is the

most common blood type.

• Individual with Rh+ if
• Rh+ and Rh+
• Rh+ and Rh- (Rh+ is dominant over Rh-)

• Anti-Rh antibodies of the system are not normally present in the

plasma, but can be produced if an individual with
Rh - is exposed to Rh+ 17
•Erythroblastosis fetalis
• Rh– mother and Rh+ father  Rh+ child 1st pregnancy

• during pregnancy fetal Rh pos rbc’s escape into

maternal circulation (antigents)
• Mother develops antibodies to baby’s Rh+ antigens

• 2nd pregnancy with Rh+ child  mother’s anti-Rh

antibodies attack unborn child’s RBCs
• Prevention: shot of RhoGAM shortly after birth of first
Rh+ baby to block development of antibodies
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Treatment of Hemolytic disease of the
newborn(HDN)
• If a woman has Rh- and gives birth to a child, or if she has a
miscarriage or abortion, she is given an injection of anti-Rh
antibodies called anti-Rh gamma globulin or RhoGAM to
prevent HDN.

• The antibodies bind to the fetal Rh antigens and inactivates

them if they crossed the placenta during birth, and the
mother’s immune system does not respond by producing
antibodies.

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Slide Blood Typing
• Very rudimentary method for determining blood
groups.
• CANNOT be used for transfusion purposes as false
positives and negatives do occur.
• A “false positive” is when agglutination occurs not
because the antigen is present, but cells may
already be clumpled.
• A “false negative” is one in which the cells are not
clumped because there are too many cells or not
enough reagent.
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Slide Blood Typing -
continued
• The slide is divided into halves.
• On one side a drop of anti-A is added, this will attach to
and cause clumping of RBCS possessing the A antigen.
• On the other side a drop of anti-B is added which will
cause clumping of RBCS with the B antigen.
• A drop of RBCS is added to each side and mixed well with
the reagent.
• The slide is tilted back and forth for one minute and
observed for agglutination (clumping) of the RBCS.

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Interpretation of Slide Typing
Testing with Anti-A Anti-
Serum
• If an RBC contains the A antigen
the red blood cells will be
agglutinated by anti-A, a positive
reaction.

• If an RBC does not have the A

antigen there will be no
clumping, a negative reaction.
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 Interpretation of Slide Typing
Testing with Anti-B Anti-
Serum
• If an RBC contains the B antigen the
red blood cells will be agglutinated
by anti-B, a positive reaction.

• If an RBC does not have the B

antigen there will be no clumping
by anti-B, a negative reaction.
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Slide Blood Typing Group A
• An unknown RBC suspension is added to known anti-sera.
• The left hand of the slide contains anti-A which reacts with
the unknown cell.
• The right hand side contains anti-B which does not react
with the cell.

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Slide Blood Typing Group B
• An unknown rbc suspension is added to known anti-sera.
• The left hand of the slide contains anti-A does not react
with the unknown cell.
• The right hand side contains anti-B which reacts with the
cell.

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Slide Blood Typing Group O
• The left hand of the slide contains anti-A does not
react with the unknown cell.
• The right hand side contains anti-B does not react
with the unknown cell.

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Slide Blood Typing Group AB
• The left hand of the slide contains anti-A which
reacts with the unknown cell.
• The right hand side contains anti-B which reacts
with the unknown cell.

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Summary of Slide Typing
Anti-A Anti-B Blood Group

NEG NEG O

POS NEG A

NEG POS B

POS POS AB

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