Immunohematology & Transfusion Medicine
ARNEL G. BAYOTAS, RMT, MD, DPSP, ASCPi (MLS),
MHA
Objectives: Principles of hematology, basic concepts,
and wll also to know the guidelines in medicine including
the bank blood testing such Rh typing and ABO typing,
incompatibility testing, cross matching, and the antbody
panel and screening
IMMUNOHEMATOLOGY
• merges aspects of hematology, immunology &
genetics
• serologic, genetic, biochemical and molecular study of
antigens associated with membrane structures on the
cellular constituents of the blood
• immunologic reactions involving all blood components
and constituents
IMMUNOLOGIC PRINCIPLES
• primary immunological components: antigens &
antibodies provides basis for blood bank testing and
reactions
• CARDINAL RULE IN BLOOD BANK:
– The antigens are found on the surface of RED
BLOOD CELLS and the antibodies are found in
SERUM OR PLASMA
Immunologic reactions in immunology involves all blood
components and its constituents including your plasma,
red blood cell and platelets and wbc
We have 2 Major reactants in your primary
immunological reactions. This involves now reaction of
antigen and antibody which provides now the basis of
your blood bank testing and reactions such Rh typing,
ABO typing and cross matching
REMEMBER:
ANTIGENS – in RBC
ANTIBODIES – in SERUM or PLASMA
IMMUNOLOGIC PRINCIPLES
Antigen Antibody
A substance that on A gamma-globulin product
introduction give rise to a of an immune response,
formation of antibody that (also called
reacts specifically with the immunoglobulin) that
same antigenic substance. reacts with antigen against
which it is stimulated.
Antigen and antibody are 2 major components of
immunologic reactions in a blood banking
ANTIGEN is a.k.a AGGLUTINOGEN or
ISOAGGLUTINOGEN reacts with the same antigenic
substance
ANTIBODY is a.k.a IMMUNOGLOBULIN or
AGGLUTININ
These are your gamma globulin products of an immune
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response that reacts with your antigen that against
stimulated. The reaction of antibody and antigen should
be specific just like your enzymes and substrate in
biochemistry
IMMUNOLOGIC PRINCIPLES: Characteristics of Antigen
• Chemical structure of antigen
– Antigens composed of oligosaccharides tend to
stimulate IgM type of antibody production
– Antigens which are primarily protein in nature,
produce IgG antibody
• Degree of foreignness
• Number of antigens introduced
– Higher the dose, greater the antibody production
• Route of administration
– IV and intraperitoneal routes are more potent than
Intramuscular or subcutaneous route
In order for the antigen to be immunogenic, we have
major characteristics of the antigen
1. CHEMICAL STRUCTURE OF ANTIGEN, it s said that
is composed mainly of oligosaccharide tend to
stimulate the ABO antibody production whereas those
antigens containing protein able to stimulate IgG
antibodies.
2. chemical structure that determines the immunogenicity
and antigenicity of antigen is the DEGREE OF
FOREIGNESS.
“The greater the degree of antigenic determinant
is recognized as non-self by individual immune
system, the more antigenic it is.”
THE MORE FOREIGN = THE MORE ANTIGENIC
3. NUMBER OFANTIGEN introduced in the patient to
elicit antibody production.
THE HIGHER THE DOSE = THE HIGHER THE
ANTIBODY PRODUCTION
4. ROUTE OF ADMINISTRATION including
INTRAPERITONEAL OFFERS MORE STRONGER
STIMULUSOF YOUR IMMUNE RESPONSE TO
PRODUCE ANTIBODY than the subcutaneous or
intramuscular route
IMMUNOLOGIC PRINCIPLES
• Immunogenicity – Ability of an antigen to stimulate the
production of its corresponding antibody in a person who
lacks the antigen
– Rh D most potent antigenic
– Fya is poor antigenic
In blood banking we have so many blood group system,
we have an ABO blood group, Rh blood group, Kell,
Duffy, Kidd, Lewis, SS, MNS, P. The MOST POTENT
antigenic blood group is belong to your Rh blood
groups.
And then your DUFFY which Fya is considered as a
POOR ANTIGENIC blood groups.
We have different subgroups belongs to your duffy, we
have your duffy A positive, duffy B positive, duffy
negative Aand duffy negative B. Those patients who
adapt negative A or B are MORE RESISTANT TO
MALARIA because duffy serve as receptors of malarial
parasite to invade blood cells
IgM is bigger structure because it is pentameric but IgG
can cross the placenta but not the IgM, IgM is more
efficient and fixing your complement over your IgG
IgG is subdivided n 4 classes: IgG1,2,3 & 4
IgG1, IgG3 & IgG4 CAN CROSS THE PLACENTA
EXCEPT IgG2

Rh with D antigen are the most potent among the IgG is most abundant in serum, only cross the placenta
blood groups IgM has the highest molecular weight because it is
pentameric

IMMUNOLOGIC PRINCIPLES
• Classification of Blood Group Antibodies:
Alloantibodies Autoantibodies
– Reacts with foreign Ag Reacts with an Ag on
not present on patient’s patient’s own cells & with
own RBC that same Ag on the cells
– Most produced as result of other individuals
of immune stimulation via
transfusion or pregnancy
(usually during delivery)

Alloantibodies are antibodies that form because of

Antibodies are more proteins in nature. This antibodies foreign antigen. For example: Natransfuse ka for several
in response to antigen and the reaction is more specific. units, massive transfusion later you will develop
However, when you treat these antibodies with a certain antibodies against that unit of blood component being
enzymes with your pepsin, papain. Papain and Pepsin transfused.
produce 2 antigen binding fragment which is a 2 Fab and
1fragment of Fc (crystallisable fragment). Autoantibodies reacts with antigen on patient’s own cells
& with that same Ag on the cells of other individuals.
Immunoglobulins
The significance of this alloantibody and autoantibody, it
 Five types: IgG – IgM – IgA - IgD - IgE could affect later on the result of compatibility testing and
 Blood group antibodies are mainly confined to IgG & even discrepancy on ABO typing.
IgM.
 IgG is a smaller molecule, with a pair each of heavy &
light chains of amino acids.
 IgG antibody can just coat but not agglutinate the cells.
 IgM has 5 such pairs joined together by the J chain.
 IgM antibody agglutinate the cells bearing
corresponding antigen.
IgG are more reactive in WARM temperature,
IgM are more reactive in COLD temperature.
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All human blood type are pareho sabi until 1901 Karl Kaya nga ang pasyenteng type O, walang antigen A
Lansteiner discovered the ABO blood group by mixing and antigen B but YOU HAVE YOUR ANTIBODY
the human blood in a test tubes with other specimen of (antibody A and B)
blood, this now will result to agglutination, so by
incubating this, some individual red blood cell with the In type O, you have H-antigen but no A&B antigen
serum with other person, it may leads to agglutination
because of this different pattern of agglutination, this
may lead to discovery in identification of 3 blood types.

O originally describe as type C, this type C is designated

as type zero signifying the lack of A and B antigen.

ABO BLOOD GROUP SYSTEM

• single most important blood group for the selection and
transfusion of blood
• widely expressed in tissues & body fluids including red
cells, platelets & endothelial cells
• three antigens: A, B, H
• two major antibodies: anti-A and anti-B
• four phenotypes: A, B, AB, O → A & B Ag’s autosomal
codominant (expressed on grp A, B and AB red cells; O Major precursor of ABO antigen is the H antigen.
phenotype autosomal recessive (most frequent)
A, AB, B- autosomal dominant A and B antigen present in people of all common blood
O- autosomal recessive type. These type 1 and type 2 CHO are main
precursor substance to become H antigen by the
Dominant means possible expression of product of action of your se gene in secretions, particurlarly in
particular gene
tears and saliva including the plasma.
For example. When an individual possess gene A and
The predominant form of H antigen or precursor of H
gene B , it will now produce the blood types A and B
blood types over your gene O antigen CHO is usually type 1. This type 1 CHO is
converted to H antigen by fucosyl transferase that
For example a child receive a gene A and O, it will now facilitates the process of fucosylation which transfer the
belong to type A because type A is more dominant monosaccharide sugar to become other precursor
compare to your O blood type. substance and it controlled by se gene.

The role of se gene directly control the presence or

absence of A, B, and H antigen in the secretions.

Kaya nga kapag nadetect mo ang A, B and H antigen

secretions you are secretors are controlled by your se
gene. Positive A and positive ka sa antigen B and
antigen H that is why you are called as secretors.

Whereas in the surface of the antigen, you have also

Antigens found in soluble form, plasma and other body
fluids especially in people called secretors
For example: Paano mo masasabiing secretors ang
isang individual if this ABO antigen are found in
secretions, tears and in the saliva.
ABO antigens can be acquired
These 3 genes determines the presence or absence of
A & B but does not directly influence the production of A
& B antigen such as the A, B and O genes.
antigen A, B and H but at this point, the major CHO
which is converted to your H antigen is your type 2. This
type 2 CHO is converted to your H antigen via your
fucosyl transferase but transferred the monosaccharide
of sugars to become the H antigen and that presence of
H antigen in the surface of the RBC is because of your
control of H gene product.

A&B genes –produce detectable product while O is

amorphic or silent so it does not produce detectable
antigen or detectable product.
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The Role of H-Gene in the Expression of ABO Genes
•Inheritance of A and B genes → results in the expression
of A and B gene products (antigens) on erythrocytes, but
are NOT the direct products of ABH genes
•Each gene codes for the production of a specific
transferase enzyme → catalyzes the transfer of a
monosaccharide molecule from a donor substance to the
precursor substance
Take note: The H, A and B antigen are not the direct
product of your H, A, B gene. Kahit may H, B and
gene allele ka, hindi ka pa rin makakapagproduce H, B
and A antigen. It is not the direct product.
Kailangan mo ng enzymatic action in order to have the H
antigen which is the precursor of all ABO blood
group antigen. Each of this gene nageencode ng
enzyme like
for example: In order to produce the H antigen as the
product of H gene, you must have an important enzyme,
that is fucosyl transferase. These enzymes catalyzes
the transfer of monosaccharide molecule from a donor
substance to a precursor substance. Yun ang function
ng enzyme na yun. Itratransfer niya ang
monosaccharide molecule either the type 1 or type 2
CHO of H antigen in order to convert.
O is a silent, amorph, it does not affect the production of
A and B substance. Kaya wala siyang enzymes. Kapag
hindi naexpressed ang A and B antigen, that is type O.
There are 3 Allele genes that encodes for the expression
of A, B O. Individuals is inherited the 2 genes coming
from your parents. These genes determine the ABO
antigen present in the persons red blood cells.
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Illustration: H gene encodes for an enzyme which
converts this precursor substance in the red blood cell
into the substance which is the H antigen.
Later on ang H substance magiging precursor siya ng
A antigen and B antigen. The A and B genes encodes
the important enzymes.
In some instances, some substances are unconverted
or H substance is partially converted so the O genes
here. The O genes now encode, the active enzymes,
wala naman kasi siyang enzymes na ininencode diba.
This will now result to no conversion of the substance
to the Group O red cells.
Take note: Kaya nga, the type O , wala siyang
antigen A and antigen B pero meron siyang
antibody A and B pero meron siyang H antigen.
In some cases, a person do not inherit H gene, wala
siyang H gene and because they don't have H gene,
they are unable to produce the H substance. Therefore
even when the A and B genes are inherited, so the A
and B cannot be form now. There are individuals do
not have H gene is known as the Bombay group.,
so they are unable to produce A and B antigen.
Individuals with rare Bombay phenotype do not
expressed the H antigen on their Red blood cells.
Kung wala siyang H antigen, meron siyang H antibody.
Even the bombay phenotype(O type)kapag
natransfuse mo siya ng type O din na donors magiging
incompatible pa rin sila because the O blood type
donor has H antigen while the Bombay considered as
type O meron anti-H.
Whereas the O gene, they are silent or amorphic since it
does not appear to control the development of the
antigen of the surface of the red blood cells. Kaya nga
kapag nagblood typing ka using your type O red blood
cell using your typing sera hindi siya nagagglutinate
in your direct/forward typing kasi nga walang siyang
antigen on the surface of red blood cell(A and B antigen)

By means of these 3 allelic genes, therefore, A allelic

genes gives, gives type to A blood types whereas B
allelic genes gives rise to type B blood types. This A and
B blood types considered dominant over your O blood
types. O blood groups is the resulting inheriting neither A
or B genes(Wala siyang A or B genes), only produced H
antigen.

Example for the with AA genotypes or AO genotypes
have the blood type of A whereas with individuals with
BB genotype or BO genotypes can produce type B blood
type. Whereas AB people expressed a special dominant
meaning they are codominant, meaning they can only
give AB genotypes later on the AB blood types. AB
codominance, which means the type A or B parents can
have only the AB child. Kung type O yan hindi ko alam
parents mo.
To illustrate the mode of inheritance, using a Punnett
square, which can predict the AB blood type. For
example, a particular mating, which the group A mother
mates with group A father therefore there are 2
possibilities of genotypes the AO or AA. 50% of blood
type A or 50% of blood type O. Hindi siya pwedeng
maging AB, hindi na natin alam sino tatay mo. Unless
your father is type AB and your mother is type A possible
blood type A, B and AB.

When we say phenotype, composed only those traits or

antigen that can be directly type. Genotype is the sum of
all genes of a person that has inherited within a blood
group system.
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-using Mendellian inheritance.
Take note the MOST COMMON SUBGROUPS AMONG
THE ABO BLOOD GROUP IS THE A (A1)
SUBGROUPS. Reason for these subgroups of the
decrease amounts on the rbc
A subgroups at may subgroup A pa, because kailangan
din maexpressed ang H chain which is converted to your
A antigen
How to determine subgroups by using Lectin.
Put O blood type after H substance. Ang
pinakamaraming H substance ay ang blood type O
using your Ulex europeus or the Lectin H.
For example: Ikaw type A ka na, bakit may agglutination
doon or may incompatibility sa ABO or cross matching,
hindi pa rin kayo compatible kahit type A ka at type A
ang donors so sa ABO type A ka but sa reverse may
incompatibility or discrepancy because baka may
subgroups ka.
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For example: A1 ka na subgroups, tapos mag
agglutinate positive ka using dilichos bifluorus(Lectin
A1) pero kapag A2 subgroups ka negative ka using
your Lectin pero type A ka. The lectin H reacts on his
strongest with the O cells. O cells yung pinakamaraming
substance among the ABO blood groups.
Use Lectin para madifferentiate mo ang A1 and A2.
SUMMARY
DIFFERENTIATING SUBGROUPS OF A:
Use the following steps to help differentiate the subgroups
of A:
•Use lectin-A1 to differentiate A1 cells from all others -
will agglutinate only A1 cells
•Look for weaker or mixed field reactions
•Look for anti-A1 in serum (serum reacts with A1 cells but
not A2 cells)
•Look at strength of reactions with anti-A,B or with lectin-H
BOMBAY BLOOD GROUP:
•The Bombay blood group lacks H gene and therefore
cannot make H antigen (H substance).
•Since the H substance is the precursor for the A and
B antigens, these antigens also are not made.
•The cells type as O and the serum has anti-A, anti-B,
and anti-H since the individual lacks all of these antigens.
Anti-H agglutinates O cells.
•The only cells Bombay individuals do not agglutinate
are from other Bombay blood people since they lack
the H antigen
ABO Antibodies are natural occuring antibodies and after the addition of your anti-human globulin, no
they are produced by environmental exposure. agglutination present, it is interpreted as Rh negative
Newborns technically, they do not have ABO antibodies after the weak D testing
initially, they begin to produce antibodies detectable only
at 6 months. 2 months or 3 months, actually blood type
pa yan ng mother, hindi pa yan sa kanya.

TRUE BLOOD TYPE: at 6 months

Group A &B are IgM reacts at room temp in your cross

matching or incompatibility testing(composed of 3
phases: immediate spin, 37 degrees phase, and anti-
human globulin .

If the blood type is A or B, since they are IgM, they

could be detected using your immediate spin.
CLASSIFICATION/NOMENCLATURE SYSTEM
IgG specially in the blood group O types and IgG act on •Fischer & Race
warm temperature or 37 degrees celcius. •Three alleles: D/d, C/c and E/e
•Five antigens: D, C, E, c, e
WHAT TO DO? •d ->no D locus ->no antigenic products
1.Wash cells with saline 3-4x and repeat all tests and test for •Rosenfeld
antibodies •Numerical system
2.Test for subgroups of A using anti-A1 and anti-A •Rh1 to Rh5
3.Use cell panels to detect the specificity of abnormal Fischer &Race proposed a different inheritance theory
antibodies and the nomenclature is based on the genetic evidence
Possible investigations that you are going to take if you of the anti-allelic nature of your Rh antigen such as your
have ABO discrepancy during your ABO blood typing: D/d, C/c and E/e. If has no D is known as the Rh
Wash your blood cells, test the recipient and donor then negative.
you wash it and then incubate.

RH SYSTEM

Rh Genetics
•3 closely linked genes control the expression of ALL Rh
Rh blood group is the first and most clinically important. antigens (codominant alleles)
Rh composed of D antigen. When they agglutinate –RHD gene – determines the expression of the D antigen
–RHCE gene – determines the expression of the C,c, E,
they are Rh positive and if they didn't agglutinate they
and e antigens
are Rh negative. Kapag positive sa D antigen, they are considered Rh
positive, No D antigen=Rh negative
Before you report Rh negative, but before you report that
you have to perform weak D testing. What you have to
do is to incubate 15-30 mins and then you add the
antihuman globulin and then you centrifuge. If is positive

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Importance of the Rh System
•After the A and B antigens, the D antigen is the most
important red cell antigen in blood banking
•The D antibody can cause transfusion reactions and
hemolytic disease of the newborn
(HDN)/Erythroblastosis fetalis
•Medico-legal parenthood studies
D antigen is the most important red cell antigen in
blood banking because the antibody in the Rh can
cause the transfusion reactions and haemolytic disease
of the newborn. The D antigen of your Rh system able to
elicit/stimulate the production of D antibody.

Erythroblastosis fetalis can be caused by immune or

non-immune causes.

One of the most non-immune cause is cardiovascular

collapse, anemia (especially thalassemia).

Immune causes is due to ABO incompatibility(it

happens when the mother is type ) and the child is type Example: If the patient is a really weak du due to D-
A or B) and Rh incompatibility (usually the MOTHER deletion, you have to write D-(D negative). Meaning no
is Rh NEGATIVE and the fetus is RH POSITIVE. reactions.

For example: The donor is Rh negative meaning wala

siyang D antigen. Kapag Rh positive may D antigen.
Weak D Phenotype
•Some D-positive RBCs DO NOT react at immediate Spin
using commercial anti-D
•In these cases, AHG testing is needed to determine the
D status
Kapag Rh typing kukuha ka lang ng patient’s red cells
and then you add the typing sera. So kapag may
AGGLUTINATION, it is POSITIVE. Meaning may nadetect
na D antigen in the surface after addition of anti-D
typing sera, meaning the patient is Rh positive.
Another reason for weak phenotype is Rh null, meaning
yung red cells wala talaga siyang Rh antigen sites.
Because of this lack antigen, it causes deformity on the
cell membrane structure. Kaya nga in your PBS makikita
mo kung minsan stomatocytes baka nga yun ang
causing stomatocytsis because of the Rh null kasi
walang antigen at walang D antigen doon resulting to
abnormal appearance of RBC or it could lead to
haemolytic anemia.

Weak D (Du) Phenotype

Weak D can be inherited in three ways:
•Incomplete/Partial antigen (D mosaic)
•Due to the position effect (DELETION) AABB Standards stands for American Association of
•Weakened expression of D Blood bank standards.

ALL DONOR RED CELLS that DO NOT

AGGLUTINATE at the immediate spin REQUIRES
WEAK TESTING.

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 HEMOLYTIC DISEASE OF THE NEWBORN
What is HDN?
 Destruction of the RBCs of the fetus and newborn by
antibodies produced by the mother
 Only IgG antibodies are involved because it can
cross the placenta (not IgA or IgM)

ABO is naturally occurring

Rh antibodies are NOT NATURALLY OCCURING
Rh antibodies are implicated with
HDN(Erythroblastosis fetalis)

Rh incompatibility, remember this HDN occur when the

fetus is positive for the antigen, the antigen of the fetus
will stimulate the production of maternal antibodies.
These maternal antibodies will destroy the fetal RBC.

Cold agglutinins- these are your cold antibodies

ABO incompatibility is most common than Rh and it is

less severe. Usually the ABO HDN occurs in the mother
type O, so meron siyang antibodies A and antibodies B
against the A antigen of the child blood type na A or the
B antigen.
Should use <3 days red cells or freshly collected red
cells
Red cell suspension should be 2-5 %
Visible result of your antigen-antibody reaction as the
basis of blood typing is AGGLUTINATION OR
HEMOLYSIS
Typing sera is expired

Antibody Characteristics
•Generally IgG (except anti-E)
•React stronger (show dosage) when antigens are
homozygous (C+c-) vs heterozygous )C+c+)
•Warm reacting
–Best at AHG
–Occasionally at 37o phase
–Rarely at IS (room temp) ABO HDN occurs in type O kaya meron siyang antibody A
•Clinically significant and antibody B against to the child's blood type.
–Because they react at body temp.
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ABO HDN can occur during your 1st pregnancy because
prior sensitization is not necessary in contrast with your Rh,
na yung 1st pregnancy, hindi pa affected yung fetus but
there is prior sentization of your maternal immune system by
the fetal rbc with the Rh positive
Mas early ang signs ng hyperbilirubinemia and jaundice
sa ABO HDN incompatibility compared to Rh kasi sa 1st
pregnancy, walang pang sensitization konting hemolysis but
the evidence of jaundice and hemolysis hindi pa gaanong
evident compared to your ABO incompatibility
Increased reticulocyte because of increased demands of
hemolytic anemia kasi mabilis ang turnover. Mabilis ang
pagkasira ng red blood cell, hindi man siya umabot sa
kanyang maturity na 120 days, nasisira because of this ABO
incompatibility as responds of that to hemolysis of your red
blood cell sa bone marrow release more reticulocyte.
Increased bilirubin because of hemolytic anemia. Diba
end product ng hemolysis ng hemoglobin ay bilirubin.
Bilirubinemia type 1, B1 or the B2
Rh HDN IS THE MOST SEVERE
This Bleeding, there is accesed now of your fetal rh positive
red cell to the maternal circulation. There will be initial or
prior sensitization of your maternal immune system.
However, subsequent offspring that are D positive will be
affected now on the second pregnancy.
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Illustration showing the Rh negative mother and the Rh
positive father with Rh positive fetus.
Pathogenesis
•Maternal IgG attaches to antigens on fetal cells
–Sensitized cells are removed by macrophages in
spleen
–Destruction depends on antibody titer and number of
antigen sites
–IgG has half-life of 25 days, so the condition can
range from days to weeks
•RBC destruction and anemia cause bone marrow to
release eryhtroblasts hence the name eryhthroblastosis
fetalis)
Pathogenesis
Dito may makikita kang immature precursor cells ng RBC
doon sa peripheral blood smear, mataas ang reticulocyte
count so because of the massive released of the
precursor of the erythroblast, the ABO incompatibility is
also known as erythroblastosis fetalis.
Liver is important in protein production because of your
protein depletion will result now to edema,
hypoproteinemia and this will leads to cardiac failure now
called as HYDROPS FETALIS.
Bilirubin
 Hemoglobin is metabolized to bilirubin
-Before birth, “indirect” bilirubin is transported across
placenta and conjugated in maternal liver (“direct”)
where it isexcreted
-After birth, the newborn liver is unable to conjugate
the bilirubin
 Unconjugated(“indirect”) bilirubin can reach
toxic levels (18-20 mg/dL)
 This is called Kernicterus and can lead to
permanent brain damage
 Red Cell Serological Techniques
 ABO and Rh (D) typing
 Weak ‘D’ or Du testing
 Antibody screening & identification
 Pre-transfusion testing
 Indirect antiglobulin test
 Direct antiglobulin test
In blood banking to prevent error and blood transfusion
reactions with use of serological techniques like ABO typing
in both donors and patient will perfom also weak du testing
especially for those donors that are negative in routine rh
typing.

Rh immunoglobulin(Rhogam) ang bibigyan ay ang Antibody screening and identification panel if the cross
MOTHER hindi ang fetus matching or incompatibility in the ABO cannot be resolved.

Rhogam attaches to the fetal rbc in the maternal circulation Pretransfusion to prevent further blood transfusion
naging access na ang fetal rbc na rh positive napunta na sa
maternal circulation kasi kapag binigyan mo siya ng rhogam
ang immunoglobulin ng rh attaches to fetal RBC na present
sa maternal circulation and they are removed by the
MATERNAL SPLEEN.

Cause of the DAT is during gestation nabigyan ng rhogam

ang mother o kaya during delivery ang fetus positive for
Direct immunoglobulin test.

ABO typing can be forward or reverse.

For example in forward typing, if your blood type is A, so

you have antigen A and antibodies B, so upon addition of
anti-A typing sera na color blue, anti-B typing sera is
yellow, so kapag nagpositive ka dun anti-A typing sera na
kulay blue and negative sa anti-B na color yellow, meaning
your blood type is A.

Kapag meron ng anti-D ang mother useless na magbigay You have antigen A reacts with anti-A typing sera.
ng Rhogam.
Blue typing sera for blood type A
Yellow typing sera for blood type B
Colorless typing sera for anti-D

Sa reverse- dinedetect mo yung antibody by using

patient's serum, plasma against the known A cells and
Known B red blood cells. So alam mo na na may A cells
ka at B cells ka irereact mo dun sa patient's serum or plasma
to detect the antibody.

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 Ang antibodies present sa SERUM OR PLASMA, ang
antigen sa RBC.

Forward typing, you have to test the RBC of your patient

which is unknown, kasing ang dinedetect mo dito ay
antigen for agglutination and made to react with you typing
sera anti-A and anti-B.

sa forward typing, kung ang blood type mo ay O, meaning to

say wala kang antigen pero may antibodies kang A and B,
so meaning wala kang reaction sa typing sera in anti-A and
anti-B. Sa reverse positive yun sa known A cells and B cells,
may reaction at may agglutination.

Take note: Ang detection ng A antigen and antibodies

reactions sa blood banking either hemolysis or agglutination.

If your blood type is AB and the reverse blood typing wala ka Weak antigen is common sa matanda. Kaya sa forward
dun agglutination in both known A cells and known B cells. type A siya pero sa reverse type O or AB siya.
bakit wala ka dun antibody? Kasi ang dinedetect ng reverse
ay antibody.

So kung type O ka, meaning wala kang antigen A and B

pero meron kang antibody A and B. sa reverse typing ang
Type O ay positive in both known A and B cells na meron
siyang antibody sa serum na nagreact dun sa known A and
B cells.

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 the cell panel to detect the specificity of a normal
antibodies.

•WHAT TO DO?
1.Wash cells with saline 3-4x and repeat all tests and test for
antibodies
2.Test for subgroups of A using anti-A1 and anti-A
3.Use cell panels to detect the specificity of abnormal
antibodies

A1- eto yung subrgroups, kaya natin dinedetermine ito para

malaman if may ABO discrepancies in your forward and
reverse typing

Unexpected alloantibodies-kapag may previous

transfusion

The results of your forward and reverse must agree before

reporting out the blood type. Kasi kapag nareport mo na
yun, bibigyan ka na kaagad ng blood, so kapag trinasfuse
patay na kaagad ang pasyente.

Kapag hindi nagagree ang forward and reverse, you have to

identify the cause and trouble what is the possibilityof ABO
discrepancy.

If you really cannot resolve the problem, “kahit ginawa mo

na ang lahat”, you must report the blood type as
UNKNOWN.

Kapag may emergency, despite na hindi mo pa rin alam ang

blood type, what you are going to do ?is you have to give
now the O blood type PACKED RED BLOOD CELL Rh
Weak O agglutinins, kung group O yun sana, this is the negative especially in child bearing mothers or patients.
foward and this is the reverse, kung type O yun dapat
supposedly dapat wala syang reaction pero dito dapat may
reaction yun, pero lahat ng reverse at forward ay walang
reaction, the reason of that is missing antigen or it is weak
antibody. Kasi kung type O siya dapat positive siya sa A1
and B cells.

What are you going to do?

 You have to incubate further for 30 mins on your
suspension at room temperature. If it is still
negative, you have to incubate further at 4 degrees
celcius.
 You have also to wash the red cells and prepare
again your red cell suspension and then repeat all
the test and you have also need to test the
subgroups(test for incompatibilities) so you need to
use Lectin by Dolichos bifluorus or lectin A1or use
13 |Immunohematology Dr. Bayotas trans by: KTRC Batch 2019
 Indirect after centrifications, if it is still negative, you have
to further incubate. After you incubate, if is still negative, so
you have to report as negative. If positive report as positive.

Direct detects the sensitizations of your RBC by

immunoglobulin( IgG) whereas in indirect it detects the
vitro sensitization of RBC by your IgG
Direct and Indirect anti-immune globulin testing
Direct anti-immune globulin testing in patient’s red cells
TAKE NOTE: Recommended specimen for direct is ETDA
and then add the anti-human globulin to detect the
and for indirect is Serum, plasma or Red blood cells.
presence of your ALLOANTIBODIES. You add your anti-
human globulin then centrifuge and look for agglutination.
Kapag may weak du testing request for Indirect
If it is positive for agglutination meaning it is positive for
DAT.

DAT usually detects the possible cause of antibodies that

may result to antibody reaction.

Indirect is important in red cell in detecting the possible

isoagglutinin and autoantibodies or had a previous blood
transfusions.

14 |Immunohematology Dr. Bayotas trans by: KTRC Batch 2019

Allogeneic blood donors the requirement for hgb should
be >12.5 g/dl but for autologous, it should be 11. Pwede din makadonate ang pregnant pero after 6 weeks
pa after delivery.
Meaning ng autologous, magdodonate ka ng sarili mo and
then , gagamitin mo siya later on. If nakareceive ka ng previous blood transfusion, pwede ka
pa rin makadonate, after 1 year.
Allogeneic hct, it should be 0.38 or 38% whereas in your
autologous, 33 % or 0.33 Kung nakareceive ka ng vaccine. For example: Live
attenuated vaccine, 2-4 weeks deferral after the last dose.
Requirements of Allogeneic Donor Qualification
Kung may infectious disease ka(e.g.HIV), permanent ka ng
Drug therapy Finasteride, isotretinoin-defe 1 deferral, hind ka na pwedeng makadonate.
month after last dose
Dutasteride-defer 6 months after
last dose
Acitretin-defer 3 years after last
dose
Etretrinate- defer indefinitely
Bovine insulin manufactured in
the United Kingdom-defer
indefinitely
Ingestion of medications that
irreversibly inhibit platelet
function(aspirin) within 36 hours
of donation precludes use of
donor as sole source of platelets
General medical Free of major organ disease,
history cancer, abnormal bleeding
tendency
Family history of CJD or recipient Pwede bang makadonate ang may gonorrhea, pwede pero
of dura mater or human pituitary
after 12 months pa.
growth hormone- defer indefinitely
Pregnancy Defer for 6 weeks
Recipient of blood Defer for 12 months If patient with history of sexual contact in individual
transfusion or persons with hepatitis after 12 months pa from the last
tissue transplant contact.
Vaccinations and Recipent of toxoid, synthetic or
immunizations killed viral, bacterial, or other Kapag nagkamalaria, pweden pa pero after 3 years. Kapag
vaccine-no deferral
Recipient of life attenuated viral or hindi ka naman nagkamalaria after mong nagpunta sa
bacterial vaccine- 2- or 4-week endemic places and nagtake ng prophylaxis- after 1 year
deferral from the last dose of your prophylactic agent.
Smallpox vaccine-refer to current
FDA guidance
Other vaccines including
unlicensed vaccines-12-month
deferral
Infectious diseases- Viral hepatitis after 11th bday
indefinite deferral Positive test for hepatitis B
surface antigen
Repeat reactive test for anti-HBc
on more than one occasion
Clinical or laboratory evidence of
HCV, HTLV, or HV infection by
current FDA regulation
History of babesiosis or Chaga’s
disease
Stigma of parenteral drug use
Injection of non-prescription drugs
Risk of vCJD according to current
FDA guidelines
15 |Immunohematology Dr. Bayotas trans by: KTRC Batch 2019
Volume of blood collected: 450 ml + 10% and the 5 SERLOGICAL DISEASE na tinetest for blood donors.
maximum volume, it should be 10.5 ml/kg per donor. Kaya 1. Syphilis
dapat ang weight ng donor ay 45kgs. 2. Malaria
3. HIV
Time of collection should be less than 10 mins. Kapag more 4. Hepatitis B
than 10 mins, hindi na siya suitable for platelet 5. Hepatitis C
preparation.
Zika virus can be transmitted by blood transfusion, sexual
Collection of blood can be done by automated system so contact and bites of mosquito
we called that Apheresis.

May lifespan na umaabot ng 42 days o 35 days.

In apheresis, you have to remove 1 or more blood

components on the donors and you have to return the
blood components na hindi mo na kailangan.

Kapag open system preparations, the shelf life is limited

up to 24 hours. After 24 hours, the particular components
You have to label the blood typing in the blood bag. Rh
are already expired.
typing also including your weak D. Also do screening for
syphilis.
Sa closed system, it follows the shelf life of anticoagulant.

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Blood should not be 3 days old.
BLOOD TRANSFUSION GUIDLINES
Before you issue blood, check for presence of hemolysis.

Any discoloration or hemolysis, should not be transfused.

Take Note: All transfusion decisions are clinically

judgement by the attending clinician.

Major cross matching of the recipient’s serum tested

against the donor red cell.
Minor cross matching,the recipient’s red cell are tested
against the donor serum or plasma.

17 |Immunohematology Dr. Bayotas trans by: KTRC Batch 2019

RBC should be stored at 1-6 degrees celcius.
Cryoprecipitate is a good source of Factor VIII/8

Washed RBCs for History of severe allergic reactions in Fresh frozen plama is for deficiency of coagulation factors.
order to remove the protein(Ig A)

RBCs leukocytes reduced is for history of febrile reactions

due to leukocytes

Irradiated RBC, eexpose mo yung blood unit doon sa

radiations para sa mga may Immunodeficiency,
malignancy or bonemarrow transplant.

Platelet transfusion para sa may dengue.

Sa iba, kahit 100 na lang ang platelet, 90, or 70, hindi pa rn

sila nagbibigay ng platelet concentrate kasi ang rason nila
may HEMODILUTION lang because of plasma na nandun
ang platelet.

18 |Immunohematology Dr. Bayotas trans by: KTRC Batch 2019

 3.There is potential for developing coagulopathy due to
coagulation factor and platelet consumption, dysfunction,
or dilution.

TRANSFUSION COMPLICATIONS
Suspected transfusion reaction
Unusual signs or symptoms temporally related to transfusion

HUWAG IREF ang PLATELETS, it should be at ROOM

TEMPERATURE. Life span of platelets is 5 days lang. It
should be under constant agitation.

Ang 35 days ang packed rbc

TRANSFUSION IN SPECIAL CLINICAL

CIRCUMSTANCES
Emergency release
Massive transfusion

Emergency release
•If time allows for typing, give ABO & Rh compatible;
otherwise give O-neg RBCs
•Label must indicate that xmatch wasn’t completed.
•Physician must sign emergency release. notified
immediately of any incompatibility.
Emergency release kung mataas ang mortality or morbidity
rate ng patient. During acute trauma,hypovolemic shock, the
ideal blood type id PACKED RED BLOOD CELL Rh
negative TYPE O.

Massive transfusion
Definitions:
1.The patient is transfused total blood volume (10-15
units of RBCs in a 70kg patient)
2.Half of the patient’s blood volume is replaced within 3
hours
3.>4 units of RBCs are transfused within 4 hours with
continued bleeding
Coagulopathies – the one complication of massive
transfusion

Potential complications of Massive transfusion

1.Impaired release of oxygen from hemoglobin
2.Transfusing massive amounts of blood may lower pH
and raise the potassium, lower the body temperature
and raise the free hemoglobin.

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 Iron overload especially in massive transfusion, you have
to give iron chelation. Oral ang defesirox at yung
TRALI stands for Transfusion related acute lung injury desferoxamine given as subcutaneously overnight.

Stop the drip pero huwag naman agad i unhook and then
you leave the IV line open with saline infusion.

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You must have written report within 7 days. It should be
attached in your medical chart of your patient.

21 |Immunohematology Dr. Bayotas trans by: KTRC Batch 2019