IMMUNOHEMATOLOGY &

BLOOD BANK
Presented by
Alyazeed Hussein, BSc
Outline

 Blood group inheritance.

 ABO system and ABO discrepancies.
 Rh system and Hemolytic disease of Newborn
(HDN).
 Minor blood group systems.
 Crossmatch and special tests.
 Blood donation, transfusion therapy.
 Transfusion reactions.
Blood group inheritance: ABO
 One position, or locus on each chromosome 9 is occupied by an A, B, or O gene.
 an individual inherits one ABO gene from each parent and that these two genes determine
which ABO antigens are present on the RBCs membrane.
 Mendelian genetics.
 Codominant in expression(Except O gene recessive!)
 ABO subgroups (A1 and A2):
 80% of all group A or AB are A1or A1B, while the remaining 20% are A2 or A2B.
 Differentiation of A1 and A2 phenotypes can be determined by using a reagent made from the
seeds of the plant Dolichos biflorus, which serves as a source of anti-A1. This reagent is known
as anti-A1lectin which reacts with A1 or A1B but not A2 or A2B.
Genotypes and phenotypes

Phenotypes Genotypes

A AA or AO

B BB or BO

O OO

AB AB
 A1 and A2 phenotypes

ANTIBODIES IN REAGENTS
SERUM

unexpecte common Anti-A1 Anti-A,B Anti-B Anti-A phenotype

d

none Anti-B 4+ 4+ 0 4+ A1

Anti-A1 Anti-B 0 4+ 0 4+ A2
 Formation of A, B, and H Red Cell Antigens

 ABH genes code for produce specific glycosyltransferases that add sugars to a basic precursor
substance.
 A, B, and H antigens are formed from a basic precursor material (called glycan) to which sugars are
attached in response to specific enzyme transferases.
 The H antigen is the precursor of A and B antigens.
 Inheritance of the H gene results in the formation of the H antigen, H and Se genes influence A and B
antigen expression.
 The H gene must be inherited to form the ABO antigens on the RBCs, and the Se gene must be inherited
to form the ABO antigens in secretions.
 When L-fucose sugar attaced to an oligosaccharide chain on the terminal galactose of type 2 chains H
substance is formed.
 When N-acetyl-D-galactosamine sugar transfer to the H substance the blood group A is formed.
 The blood group B is formed if D-galactose sugar transfer to the H substance.
 Genotype hh (Bombay), no production of α-2-L-fucosyltransferase no L-fucose, no A, B, O Ags
produced.
H: O > A2 > B > A2B > A1 > A1B

ANTIGEN SUGAR GLYCOSYLTRA Gene

NSFE-RASE

H L-fucose α-2-L- H
fucosyltransferase

A N-acetyl-D- α-3-N- A
galactosamine acetylgalactosami
nyltransferase
B D-galactose α-3-D- B
galactosyltransfer
ase
The Bombay Phenotype Oh (H null)
 First reported by Bhende in1952 in Bombay
(India).
 hh genotype.
 No H antigens formed; therefore, no A or B
antigens formed.
 Phenotypes as blood group O.
 Anti-A, anti-B, anti-A,B, and anti-H present
in the serum.
 Can only be transfused with blood from
another Bombay (Oh).

Glycan and RBCs

 Reverse grouping Forward grouping Reverse grouping Forward grouping

B Cells A1 Anti-B Anti-A B Cells A1 Anti-B Anti-A

Cells Cells
4+ 4+ 0 0 Patient 0 0 0 0 Patient

Group O Group !!!?

 ABO Discrepancy

 ABO discrepancies occur when unexpected reactions occur in the forward and reverse grouping.
 patient’s serum problems (reverse grouping), or patient’s red cells (forward grouping).
 The unexpected reaction can be due to an extra positive reaction or a weak or missing reaction in the
forward and reverse grouping.
 note that!! the RBC and serum grouping reactions are very strong (3+ to 4+), weaker reactions usually
represent the discrepancy.
 Group I Discrepancies (more common): Affect reverse grouping.
 Due to depressed Ab production:
 Newborn.
 Elderly patients.
 Leukemia(hypogammaglobinemia).
 ABO subgroups.
 Plasma transfusion and exchange transfusion(Dilution).
Group II Discrepancies
 Affect the forward grouping.
 ABO subgroups.
 Leukemia.
 Acquired B Ag phenomenon (colon cancer) weak react with Anti-B.
 Group III Discrepancies:
 Affect both, forward & reverse grouping.
 Protein or plasma abnormalities that cause rouleaux formation.
 Due to high globulin: Multiple myeloma or Waldenstron macroglobulinemia.
 High fibrinogen.
 Group IV Discrepancies:
 Affect the forward & reverse grouping.
 Cold reactive autoantibodies.
 RBCs other than ABO(RBCs transfusion or marrow/stem cell transplantation).
 Alloantibodies.
 Reverse forward
Possible causes Autocon Screenin
-trol g cells
B cells A1 Anti-B Anti-
cells A

Group O newborn, 0 0 0 0 0 0 1
elderly patient low
Igs.
Rouleaux, cold 2+ 2+ 2+ 2+ 4+ 4+ 2
auto-Ab
A2 subgroup(anti- 0 0 4+ 1+ 0 4+ 3
A1)
A2B subgroup(anti- 0 0 0 1+ 4+ 4+ 4
A1)
Oh (Bombay) 0 4+ 4+ 4+ 0 0 5

Acquired B 0 0 4+ 0 2+ 4+ 6
phenotype
 Agglutination reactions grading

 Zero: lowest grade; no agglutinative red

blood cells are present.
 1+: red blood cell button divides into a
number of small and medium-sized
clumps.
 2+: red blood cell button divides into
numerous medium-sized clumps.
 3+: red blood cell button divides into
large clumps.
 4+: red blood cell button does not break
into clumps, free red blood cells cannot
be seen in the background.
 Hemolysis: positive.
 Rh Blood Group System
 Rh refers to a specific red blood cell (RBC) antigen (D).
 Rh-specific antigens reside on proteins versus the carbohydrate antigens ABO and Hh.
 Rh antibodies are produced only after exposure to foreign red blood cells(HDN).
 A primary cause of hemolytic disease of the fetus and newborn (HDFN, also called
erythroblastosis fetalis).
 Five antigens made up the Rh system(D, C, c, E, e).
 RH Genes: two closely linked genes located on chromosome 1 control expression of
Rh proteins namely, RHD and RHCE.
 RHD codes for RhD protein, RHCE codes for either RhCe, RhcE, Rhce, or RhCE
proteins.
 Codominant.
 Rh terminology: Fisher-Race: DCE Terminology

 They postulated that the antigens of the

system were produced by three closely
linked sets of alleles.
 Each gene was responsible for
producing a product (or antigen) on the
RBC surface.
 D, d, C, c, and E, e, (d: absence of D
Ag).
 each person inherits a set of Rh genes
from each parent (i.e., one D or d, one C
or c, and one E or e).
 Rh phenotype is reported as DCE rather
than CDE.
 C, c, E, and e recognized by specific
antibodies.
 Wiener: Rh-Hr Terminology
 One gene responsible for defining Rh.
 R denotes the presence of the D antigen, while
the r indicates the absence of D antigen.
 Presence of C is indicated by a 1 or a single
prime (´ ), while in c no 1 or (´ ).
 Presence of E is indicated by 2 or double prime (
˝ ), While in e there no 2 or ( ˝ ).
 Presence of both C & E indicate by Rz or ry
(DCE or dCE) respectively.
 If the r precedes the h (i.e., rh ´ or rh ˝), this
refers to the C or E antigens, respectively.
 If the h precedes the r (i.e., hr ´ or hr ˝ ), this
refers to th c or e Ags, respectively.
 Rh0 = D.
 Rosenfield and Coworkers: Alphanumeric
Terminology

 Number to each antigen of the

Rh system.
 A minus sign preceding a
number designates the absence
of the antigen.
 Rh1= D, Rh2= C, Rh3= E, Rh4=
c, and Rh5= e.
 D + C+ E + c negative, e
negative = 1, 2, 3, –4, –5.
 If e not tested Rh will be: 1, 2, 3,
–4.
 Weak D

 Possess D antigen that requires an indirect antiglobulin test to detect the presence of D antigen.
RBCs carrying weaker D antigen have historically been referred to as having the Du type.
 Immunogenicity: ABO, Rh(D > c > E > C > e), Kell, Duffy, Kidd.
 Minor blood group systems
Systems with cold antibodies:
Lewis:
 Lewis antigens: Le & Le antigens.
 Soluble Ags produced by tissues, then adsorbed to RBCs surface.
 Lewis antibodies: IgM, clinically not significant.
I:
 I antigens: I (adult), i (newborn).
 I antibodies: Anti-I (IgM), Cold agglutinin disease, Secondary to mycoplasma infection.
P:
 P antigens: P1 & P2.
 P antibodies: Anti-P1: IgM, clinically insignificant. Anti-P: IgG, Paroxysmal cold hemoglobinuria(PCH), Also
called Donath Landsteiner antibody.
MNSs:
Contain ( M, N, S, s, and U Ags).
 MNSs antibodies: Anti-M & anti-N (IgM), insignificant. Anti-S, anti-s and anti-U (IgG), can cause HDN.
 Systems with warm antibodies

Kell system:
 Kell Ags: K Ag (<9%), k Ag (>90%), K Ag more immunogenic after D Ag.
 Kell Abs: Most common Anti-K (IgG), can cause HDN.
Kidd system:
 Kidd Ags: Jk(a) and Jk(b).
 Kidd Abs: Anti-JK(a) and anti-Jk(b), (IgG), cause HDN.
Duffy system:
 Duffy Ags: Fy(a) and Fy(b).
 Duffy Abs: IgG, not a common of HDN.

Null phenotypes!!!!
 :HUMAN LEUKOCYTE ANTIGENS (HLAs)
 Genes of HLAs are part of major histocompatibility complex.
 Located in chromosome 6 and is divided into Class I, II and III:
1. Class I includes A, B and C loci.
2. Class II includes DR, DP and DQ.
3. Class III includes complement proteins.
 Class-I on platelets and nucleated cells.
 Class-II on APCs-phagocytic cells (B-lymphocytes, monocyte, macrophages, neutrophils, dendritic cell) &
activated T cells.
 Contributes to self and non-self recognition.
 Immune response to transfused incompatible HLAs causes fever and chill (known as febrile non-hemolytic
transfusion reaction.
 HLA must be matched for organ, tissue, bone marrow and stem cell transplant donors and recipient, if the
recipient is not matched correctly a severe graft-versus-host disease results (GVHD).
 HLA test applications include paternity testing, organ and tissue transplantation, bone marrow and stem cell
transplantation and platelet matching.
:Transplantation

Types of graft:
 Autograft: Transfer of tissue from one site to another within an individual.
 Isograft (syngraft): Transfer of tissue between genetically identical individuals.
 Allograft: Transfer of tissue between two genetically nonidentical individuals of the same
species.
 Xenograft: Transfer of tissue between two individuals of different species.
Platelet antigens:
 Membranes have protein antigens.
 Platelet antibodies occur less frequently in the general population because of less Ag variability.
 Antibodies reacting with platelets may be ABO-HLA, or platelets specific.
 Diseases: neonatal alloimmune thrombocytopenia and posttransfusion purpura.
Antigens of high incidence: Ags that occur in at least 98% of the
population.
 You know you have an antibody to high-frequency antigen when:
 The autocontrol is negative.

 Reactions occur at AHG phase.

 I, H, P, P1 (room temperature reactions).

 Lu(b), Ch, Rg, Csa, Kna, McCa, Sia and JMH (at AHG).
Antigens of low incidence:
 the antibody screen is negative, and the crossmatch is positive.

 Lua, Cw, Kpa, Wra, V, Bga, VS and Cob.

Antibody screen:
 Purpose:
 patients requiring transfusions, pregnant women, blood and blood product donors, and patients with
suspected transfusion reactions.
 Use to detect unexpected antibodies (other than anti-A and anti-B).
Procedure
1) Mix known RBCs with patient's serum.
2) Add potentiator (Albumin, saline) to enhance agglutination and incubate at 37°C.
3) Spin and read results (if applicable to potentiator).
4) Wash three times with saline.
5) Add AHG, spin, then read results.
6) Read all negative results macroscopically (some facilities read all negative results microscopically).
7) Add IgG-coated control cells (check cells) to all tubes with a negative reaction at AHG. Check cells
must be agglutinated, or the test must be repeated.
8) Results: Any agglutination at any phase of testing indicates an atypical or unexpected antibody.
 Antibody identification: Antibody panel: Type O cells with known antigens; usually 10-20 bottles of different
cells with known antigens. Purpose: To identify alloantibodies detected in patient's serum. The reaction
phase of the antibody is important. It will determine if the antibody is IgG or IgM. Room temperature
reactions usually indicate an lgM antibody. Reactions at 37 °C and/or AHG usually indicate an IgG antibody.

 Auotoantibodies: Detected by a positive DAT or positive auotocontrol. Produced in response to drug effect,
cold autoimmune disease, pneumonia, warm autoimmune disease, infectious mononucleosis.

 Alloantibodies: Cold Abs, that react at 4°C and/or room temperature are usually not clinically significant.
These antibodies can hide a clinically significant alloantibody.
 Positive screen cells and negative autologous control.
 Prewarmed techniques or adsorption of cold antibody can help detected any alloantibodies present if the cold
antibody reacts at 37°C, it may be clinically significant.
 prewarmed technique used to prevent cold-reactive alloantibodies or autoantibodies from reacting in the IAT
phase.
Elution:
 Transfusion reactions, HDN.
 IgG that attaches to RBCs in vivo can be removed by elution (in vitro),
 3 types of elution techniques:
 Intact RBC antibody removal uses buffers to remove the Ab from the RBC without

destroying the RBC.

 Digitonin release the Ab by destroying the RBC.

 Lui freez-thaw is used to remove IgM Abs (usually A or B) present on newborn RBCs.

Adsorption:
 Adsorption refers primarily to the adherence of an antibody or antigen onto the surface of a red
blood cell, used by blood bankers to bind antibodies to red blood cells in order to remove them
from the plasma.
Autoadsorption: using the
patient’s own RBCs to remove
autoantibodies (warm
autoantibodies).
Alloadsorption: using selected
non-self RBCs to remove 
alloantibodies.
 Antiglobulin test(Coombs’ test), anti-human globulin (AHG) test

Use an antibody to find another antibody!!

 Use for detection of red cell antibodies.
 Principle: an antibody (anti-IgG) to gamma globulin can form bridges between red cells
sensitized with antibody (incomplete Ab) and cause them to agglutinate(complete agglutination).
 2 types: direct anti-globulin( DAT) and indirect anti-globulin test (IAT).
Direct Antiglobulin Test (DAT, DCT, DAGT):
 Detects antibody in vivo sensitizing RBCs.
 Uses patient cells.
Indirect Antiglobulin Test (IAT, IDC, IAGT):
 Detects free antibody.
 Uses incubation at 37 ºC to force red cell sensitization in vitro.
 Uses patient serum.
 Direct Antiglobulin Test (DAT, DCT, DAGT):

1. One drop of patient RBCs are washed with 0.9% NaCl 3 times(minimum) to remove the
unbound Abs.
2. AHG reagent is added.
3. Tube is centrifuged.
4. Positive agglutination = patient cells coated with Abs.
 Application of DAT:
1. HDFN (Hemolytic disaes of the fetus and newborn).
2. AIHA (Autoimmune hemolytic anemia), cold agglutinin disease(DAT positive = complement),
warm auto immune hemolytic anemia (DAT positive =IgG and C3).
3. Drug-related hemolytic anemias.
4. hemolytic transfusion reactions.
 Indirect Antiglobulin Test (IAT, IDC, IAGT):

1. One or two drops of serum is added to test tube.

2. One drop of RBCs is added.
3. Incubation of tube at 37 ºC.
4. Wash 3 times with saline.
5. Add two drops of AHG reagent.
6. Centrifuge the tube and observe the agglutination.
Aplication of IAT:
7. Crossmatch (patient serum is incubated with donor red cells)
8. antibody detection (patient serum is incubated with group O screen cells)
9. antibody identification (patient serum is incubated with group O panel cells)
10. antigen typing (commercial antiserum is incubated with patient or donor cells).
 Crossmatch
 It’s a procedure used prior to a blood transfusion to determine whether donor blood is compatible
or incompatible with recipient blood(not be evident on blood typing!
 Types:
1. Major crossmatch: testing the patient plasma with donor cells (detect patient Abs directed
against donor cells).
2. Minor crossmatch: testing patient cells with donor plasma ( detect donor Abs directed against
patient cells).
Procedure:
3. For major cross matching: label test tube mix two drops of patient serum with one drop of
donor cells.
4. For minor cross matching: label test tube mix two drops of donor serum with one drop of
patient cells.
5. Mix the tubes and incubate at 37 ºC for 45 minutes. Add two drops of AHG and mix well.
6. Centrifuge for 1 minute at 1500 rpm. Read macroscopically and microscopically. No
agglutination: compatible. Agglutination: incompatible.
Donor selection criteria

 Age: ≥ 16 years.
 Temperature: ≤ 37.5ºC or

≤ 99.5ºF.
 Blood pressure: 80/50 mm/Hg

And not higher than:

180/100 mm/Hg
 Hb: ≥12.5g/dL, HCT: ≥ 38%.

 Weight: minimum 110 Ibs/ 50kg.

 Donor testing

HBV:
1. Positive anti-HBs > vaccination.
2. Positive for HBsAg, anti-HBc and anti-HBc(IgM) >
active infection.
3. Positive for HBsAg, anti-HBc > chronic infection.
4. Positive for anti-HBc and anti-HBs > immunized.

HDV!!(associated with HBV).

 Whole blood collection

 Whole blood contains RBCs and plasma, with a hematocrit level of approximately 38%.
 The platelets, white cells, and labile clotting factors do not survive in stored whole blood.
 It must be stored at 1°C to 6°C, and the shelf-life is dependent on the preservative used:
1. ACD or CPD: 21 days.
2. CPDA: 35 days.
3. SAGM(Saline adenine glucose manitol): 45 days.
Apheresis donation:
 Apheresis collection is an effective mechanism for collecting a specific blood component while
returning the remaining whole blood components back to the patient.
 Most apheresis instrumentation use an automated cell separator device whose centrifugal
force separates blood into components.
 Used to collect platelets, plasma, leukocytes and red cells.
Apheresis donation

Plateletpheresis.
 Plasmapheresis.

 Leukapheresis.

 RBC Pheresis.
Autologous Donors:
 The patient banks his/her own blood at least two weeks before an elective
procedure for self-use.
 Most autologous blood is used to treat surgical blood loss in very specific
situations where there is a reasonable opportunity to avoid homologous
transfusions or when compatible allogeneic blood is not available.
 Rare blood groups.

Hb ≥ 11.0 g/dL or HCT ≥ 33%.
Direct donation:
 A “directed” blood donation occurs when a patient gets the opportunity to choose
their blood donor!
 Typically, the donors are family members of the patient.
  They are NOT any safer than getting blood from volunteer, allogeneic donors!!
  Directed donors are very commonly first-time blood donors.
 Components storage and transfusion therapy
indications Storage Shelf-life Blood
temp. components
O2 ↑ 1–6°C CPD-21 d RBC
CPDA-1 35
d
CP2D-21 d
ACD-21 d
AS-42 d

Thrombocytopenia 20–24°C 5 days PLT

DIC, bleeding 4hr PLT, pooled
Coagulation –18°C 1 yr FFP
deficiency –65°C 7 yr
Liver disease
DIC
Hemophilia A, VWD FXIII deficiency, –18°C 1 yr Cryoprecipitate
Hypofibrinogenemia
Neutropenia 20–24°C 24 hr Granulocytes
Donor reactions:
 donor may develop nausea, dizziness, specially if you are collecting

whole blood or he/she is sensitive to citrate for apheresis. Stop the

collection immediately and Remove the tourniquet.
Autoimmune hemolytic anemia(AIHA):
 Destruction of RBCs by autoantibodies,

 Auto= these Abs directed against individual’s own RBCs.

1. Warm AIHA(WAIHA): IgG.

2. Paroxysmal cold hemoglobinuria(PCH).
 Transfusion reactions

 Acute(hemolytic): Occure immediately after a blood transfusion due to incompatible transfusion.

Signs and symptoms are: fever; chills; fast heart beat (tachycardia); blood clotting (DIC); bloody
urine (hemoglobinuria); low blood pressure (hypotension); kidney failure; and cardiac collapse.
 Delayed-immune mediated hemolytic: Within five to seven days after a transfusion. Patient
produce Ab against non ABO RBCs Ags (kell, kidd). Signs are fever or mild jaundice.
 Anaphylactic transfusion reaction: immediately after a transfusion is initiated, from a severe
allergic reaction. Signs are bronchospasms, wheezing, and cough, but not fever. Anaphylactic
transfusion reaction is fatal if not treated immediately. Usually, the problem is that your patient
inherited an immunoglobulin A (IgA) deficiency. Complement-binding anti-IgA antibodies form
when your patient is exposed to the donor's IgA.
 Non hemolytic febrile reaction: Cytokines form when blood is stored. Your patient may develop
a fever as a reaction to the donor's cytokines, but it is unlikely that he/she will go on to develop
breathing problems (respiratory distress) or low blood pressure (hypotension). The fever will
probably be self-limiting, but you must still notify the doctor, nurse, and lab manager, as a
precaution.
 Transfusion-related acute lung injury (TRALI): The donor's antileukocyte antibodies attack the
recipient's white blood cells, which activates complement. Blood vessels in your patient's lungs
become more permeable and leak, so your patient develops pulmonary edema, cyanosis,
hypoxia, and dyspnea.
 Graft-versus-host disease (GVHD): If the patint is immunocompromised, then the donor's
lymphocytes are not killed and GVHD results. Consider giving them irradiated blood to prevent
GVHD. Signs: fever; diarrhea; sloughing skin; chronic cough; irritated mouth and eyes; muscle
cramps; and abnormal liver function.
 Prevention
Give ABO compatible blood
Give antigen negative blood
Wash cellular component or blood
product from IgA deficient
Leukocyte reduced unit or plasma
removal
Washed RBCs from which plasma is
removed, deferring multiparous
female, male products,
leukoreduction
Irradiated blood unit
Etiology Transfusion reactions
ABO antibodies (IgM, IgG, complement Acute hemolytic
activation)
Non-ABO antibodies (Rh, Kell), Delayed immune hemolytic
complement is not activated
Recipient anti-IgA directed against donor Anaphylactic reaction
.IgA
Recipient antibodies to donor WBCs or Febrile non-hemolytic
cytokines
Donor antibodies to recipient HLA or TRALI
WBCs (neutrophils)
Donor T-lymphocyte (CD8) attack GVHD
recipient tissues
 hemolytic disease of the newborn (HDN)

 AKA Erythroblastosis fetalis, the

pregnant mother has a different blood type
than her unborn infant. The problem may
be either Rh or ABO incompatibility.
 The mother's IgG antibodies travel through
the placenta and attack the red blood cells
of her fetus. The fetus may become
anemic, leading to heart failure.
Alternatively, hemoglobin from destroyed
red blood cells is converted into indirect
bilirubin, leading to jaundice, retardation,
brain damage(kernicterus), deafness, and
sometimes perinatal death.
 The first baby not affected.
 The baby is edematous, with hydrops and a swollen liver or
.spleen

 The most severe, but least

common, form of HDN is Rh
incompatibility. A D- mother
develops D antibodies in response
to a D+ baby. If the same mother
subsequently has another D+ fetus,
then the D antibodies from her first
pregnancy attack the red blood
cells of the second fetus. The
delivery team performs an
exchange transfusion, aggressive
hydration, and phototherapy to
reduce the complications of HDN,
which are neurological syndrome,
kernicterus, hydrops fetalis, and
hypotonia.
 Rh incompatibility HDN is preventable if the mother
receives injections of RhoGAM immune globulins.
 ABO hemolytic disease is a less severe, but more
common, form of HDN. A type A mother reacts to a
type B or AB fetus; or a type B mother reacts to a type
A or AB fetus; or a type O mother reacts to an A, B, or
AB fetus. ABO hemolytic disease is treated with
phototherapy, to remove excess bilirubin.
 HDN prevention: A pregnant D- mother receives anti-D
intramuscularly between 26 and 28 weeks & within 72
hours after delivery.
Fetal screen (Rosette test): Maternal RBCs is
incubated with anti-D. Anti-D binds to Rh-
positive fetal RBCs, if present in the maternal
circulation. D-positive indicator cells are added
that bind to the anti-D, forming a rosette
around the sensitized Rh-positive fetal RBCs.

Kleihauer-Betke (KB) acid elution is used to

determine the amount of a fetomaternal
hemorrhage. Principle: Fetal hemoglobin is
resistant to acid elution. A blood smear from
the mother is made, then dipped in an acid
buffer and stained with a counterstain. The
buffer lyses the mother's cells (ghost cells) and
does nothing to the fetal cells. Pink fetal cells
are counted.
Intrauterine transfusion (IUT) and neonatal transfusion

 O Rh-negative RBCs should be

selected for the intrauterine
transfusion.
 Less than 7 days old, To reduce
the risk of hyperkalemia!!!
 Irradiated to prevent GVHD.
 Leukoreduced unit.
 Negative for CMV
(seronegative).
 Negative for Hb-S.