ABO Blood Group

Karl Landsteiner Gene

H
Glycosyltransferase

L-fucosyltransferase
Immunodominant sugar

L-fucose
Proponent of ABO blood group A N-acetylgalactosaminyltransferase N-acetylgalactosamine
First to perform ABO grouping B D-galactosyltransferase D-galactose
Observed 3 patterns of activity
A, B, and C -> A, B, and O
Antigen Antibodies
Blood group present in present in Genotype
ISBT 001- ABO Blood Group RBCs serum

Most important and most clinically significant A A Anti-B AA or AO

Found in Chromosome 9 B B Anti-A BB or BO

Arise from a single ABO gene mutation:
AB A, B None AB
Substitution
Frameshift Mutation O None Anti-A, Anti-B OO
Hh Genes - needed to form ABO antigens on
RBC membranes; Loc: Chromosome 19 Group O > Group A > Group B > Group AB
Se Genes - needed to form ABO antigens o
secretion; Loc: Chromosome 19 Amount of H antigen:
O > A2 > B > A2B > A1 > A1B
ABO Ag/Ab Detection
Blood typing: Forward and Reverse

Forward/Direct typing Reverse/Indirect typing

Specimen -> RBCs (Ag) Specimen -> Serum

Reagents -> Anti-A = Asul = Blue: Thymol blue Reagents -> Known A1 and B cells
Anti-B = Banana = yellow: Acriflavine Not recommended for Newborns and cord blood
Anti-D: colorless
Lectin Acquired Pseudo Antigens

Anti-A1 (Dilochos biflorus) Acquired "A" Antigen

Anti-B (Griffonia simplicifolia) -> type O or B caused by Proteus mirabilis
Anti-H (Ulex europaeus) Acquired "B" Antigen
-> type A and O caused by P. vulgaris and
Bombay Oh(Hnull) E. coli 086

double dose of h gene Parabombay Phenotype

hh genotype
devoid or have small amounts of H antigens
ABO
ABO DISCREPANCIES
DISCREPANCIES
Discrepancies Causes Resolutions

Check px history
GROUP I (Problem Weakly reacting/missing Incubate px serum at room temp. (15-30 mins)
in Reverse Typing) antibodies If negative rx after centrifugation = lower temp. to 4°C
(15 mins)

GROUP II (Problem Weakly reacting/missing Incubate at room temp. (30 mins)

in Forward Typing) antigens If still negative = incubate at 4°C (15 mins)

Microscopic examination
Protein/Plasma
GROUP III Saline replacement
abnormalities
Wash cells with saline (3x)

Incubate at 30°C (15 mins); Wash cells with saline (3x) &
Cold autoantibodies & non- retyped
GROUP IV
expected alloantibodies Treat with 0.01M Dithiothreitol
Run DAT, AC, and Antibody Screen
 Rh Blood Group
HISTORY Rh Gene
Levine and Stetson
RhD -> presence or absence of the RhD
1939
RhCE -> RhCe, RhcE, Rhce or RhcE
Acute Hemolytic Transfusion Reaction (AHTR)
RhAG -> a coexpressor
D positive (Rh +) -> RHD + RHCE
1940
Landsteiner and Weiner
D negative (Rh -) -> complete deletion of RHD
Discovery of Rh Blood group through the
Rhesus macaque monkey

Rh Phenotypes
Nomenclature for Rh System Rh Positive: inherits one or two RhD genes
Rh Negative: arise from 3 different mutations
Fisher-Race (genetics & serology)
European: Deletion of RhD gene
Weiner (short-hand method)
African: 66% RhD pseudogene
Rosenfield and Coworkers (presence or
Asian: alteration of RhD gene
absence of antigens)
International Society of Blood
Transfusion Committee (ISBT) (catalogue
of each antigen)
Fisher-Race Terminology (DCE Terminology)
D antigen
most clinically significant antibody in pre-transfusion testing
presence of D antigen (D+) / absence of D antigen (D-)
Cc antigen
Codominant 
Less dominant Immunogenicity
Ee antigen
Codominant D>c>E>C>e
e is more frequent than E

Rh/D Positive Rh/D Negative

Dce dce or ce

DCe dCe or Ce

DcE dcE or cE

DCE dCE or CE
 CE Fisher - Fisher -
D(+) D(-)
antigen Race Race

Ce R1 DCe r' dCe Weiner Nomenclature

cE R2 DcE r" dcE 5 major Antigens: Rh0, rh‟, rh‟‟, hr‟,hr”
Conveys the Rh haplotype
cc R0 Dce r dce

CE Rz DCE ry dCE

Alpha-numeric terminology
Rosenfield and Workers Assigns a number to each antigen of
the Rh system
International Society Of Blood Transfusion Committee (ISBT)
Machine readable
Adopted a 6 digit number for authenticated antigen belonging to the blood group system

Weak D D- ANTIGEN DETECTION

Considered as Rh positive Slide Testing
Requires IAT Tube Testing
Automated & Microplate Testing
Variant Gel Testing

C-trans to RHD (position effect) HEMOLYTIC DISEASE OF THE

Weak D (genetically transmissible)
Partial D (D mosaic) FETUS AND NEWBORN
Del Maternal antibody crosses the placental
barrier and sensitizes the fetal RBCs
Rh Deficiency Syndrome
Rh Immune Globulin (Rhogam)- given to D
Rhnull negative pregnant women following a delivery
Fails to express any Rh antigens on the RBC surface of a D-positive fetus
Golden Blood
Rhmod
mutation in RhAG
less severe than Rhnull
 Other Major Blood Groups
Significant Blood Groups Insignificant Blood Groups

Kell Lewis

Duffy P1

Kidd MN

Ss Lutheran

I/i

KELL (006)
First blood group system after the discovery of Coombs test
Kx System (019) - absence of Xk results in McLeod Syndrome (very rare infecting only males)
K and k Antigens - not denatured by ficin & papain but destroyed by trypsin & chymotrypsin
Anti - K - most common antibody seen in the bloodbank
K0 Phenotype - may produce anti-Ku if immunized
 DUFFY (008)
Named after Mr. Duffy (found to
have the first Anti-Fya)
Fyb antigen - serum of a
woman (3 pregnancies)
Fy(a-b-) - resist P.knowlesi
and P.vivax
FyFy - common genotype in
blacks
KIDD (009)
Common cause of HTRs
Jka and Jkb Antigens -
treatment of RBCs with
enzymes enhances
reactivity
Anti-Jka and Anti-Jkb -
notorious reputation
Jk (a-b-) Phenotype - lack
Jka , Jkb , Jk3
Recessive Allele, Jk -
“silent” allele Jk
LEWIS (007)
adsorbed from the plasma
Anti-Lea - Causes HTR if
transfused with LE(a+)
Anti-Leb - IgM agglutinin and
can bind complent
LUTHERAN (005)
Lewis Antigens - readily found in serum of a lupus
shed from transfused RBCs erythematosus patient
Lua and Lub Antigens -
MNS (002) poorly developed at
birth
M and N Antigens- Easily Anti-Lua - some may
destroyed bind complement
S and s Antigens - Less easily Anti-Lub - response to
degraded transfusion or
Anti-M - not clinically pregnancy
significant for transfusion Anti-Lu3 - rare
Anti-N - does not bind antibody
complement
Anti-S and Anti-s -
implicated in severe HTRs
with hemoglobinuria;
 I (027)
I antigen – cold agglutinin stand for “individuality”
i antigen – produced by a rare gene resulting to I-phenotype now known as adult i
Anti-I - M. pneumoniae stimulates the production of autoanti- I
Anti-i - associated with infectious mononucleosis (Epstein- barr virus infections)

P BLOOD GROUP
P1 Antigen – deteriorates rapidly on storage
Anti-P1 – Typically weak, cold reactive saline agglutinin
associated with Echinococcus granulosus tapeworm from P1- individuals
found in patients with fascioliasis(bovineliver fluke disease) and in bird handlers
 Minor Blood Groups
carried on band 3, a major integral
membrane glycoprotein
THE DIEGO SYSTEM (010)
useful tool in anthropologic studies of
Mongolian ancestry

located on erythrocyte
acetylcholinesterase, an enzyme
THE YY SYSTEM (011) involved in neurotransmission
Three phenotypes: Yt(a+b-), Yt(a+b+)
common, Yt(a-b+)rare

Controlled by x-linked gene

Xga antigen is carried by a protein with
cell adhesion properties
THE XG SYSTEM (012) Anti-Xg a are predominantly IgG
reactive in the IAT and sensitive to
enzymes
 Composed of three antigens Sc1, Sc2,
and Sc3 antigens
THE SCIANNA SYSTEM (013) anti-Sc1 and anti-Sc2 are implicated
with HTR and Anti-Sc3 with mild HTR

react primarily in indirect antiglobulin

THE DOMBROCK SYSTEM (014) test w/PEG or Enzyme enhancement

Composed of three antigens: Coa , Cob ,

and Co3 (formerly Coab )
THE COLTON SYSTEM (015) Channel-forming integral protein (CHIP)
- responsible for water permeability

associated with human leukocyte

antigen (HLA) system
CHIDO/ROGERS SYSTEM (017) Stimulated usually in multi-transfused
individuals

expressed on glycophorin C (GPC)

and/or glycophorin D (GPD)
THE GERBICH SYSTEM (020) present with a change in erythrocyte
morphology in the form of Elliptocytes
 regulation of complement activation by
THE CROMER SYSTEM (021) accelerating the decay of the C3 and C5
convertase

Composed of five antigens: Kna and

THE KNOPS SYSTEM (022) Knb (knops), McCa (McCoy), Sia, and
Yka (York)

carried on the hematopoietic isoform of the

THE INDIAN SYSTEM (023) CD44

THE OK SYSTEM (024) Function as receptor and adhesion molecules

from monoclonal, and Eleanor Roosevelt (lab

THE RAPH SYSTEM (025) where the antibody was produced)

THE GILL SYSTEM (029) found on the glycerol transporter aquaporin 3

Newest blood group system

THE RHAG SYSTEM (030) Absence of RhAg due to Mutations result in the
Rh null phenotype
 AHG Testing
IMMUNOGLOBULIN
also known as ANTIBODIES
Y-shaped
Light Chains - κ or λ; shorter subunit
& less dense
Heavy Chains - γ , α , µ , δ , ε; longer
and dense subunit
targets of the Antihuman Globulin
Reagent.
ANTIHUMAN GLOBULIN TESTING
also known as Coomb’s Test
ANTIHUMAN = Anti (directed against) +
Globulin (human antibodies) = Antibodies
against human antibodies
Detects the presence of incomplete or
non-agglutinating antibodies
AHG Reagent - Green
Polyclonal Antihuman Globulin
conventional method using
rabbits
Sheep, goat and horse used
when large volumes of antibody
is required
termed as Broad Spectrum
Antibody
Monoclonal Antihuman
Globulin
hybridoma technology using mice
fusing of an antibody-producing B
cell with multiple myeloma cells
Spleen cells contain HGPRT
(Hypoxanthine Guanine
Phosphoribosyl Transferase)
HAT medium= Hypoxanthine,
Aminopterin, Thymidine
Indirect Antiglobulin Test
Detects for in vitro sensitization
or coating
Specimen: Serum Sample
requires between 100 and 200
IgG or C3 molecules on the cells
to obtain a positive reaction
Direct Antiglobulin Test
Detects for in vivo sensitization or coating
Specimen: EDTA or Anticoagulated blood
samples
can detect 100 to 500 IgG molecules per
RBC and 400 to 1,100 molecules of C3d
per RBC

Clinical Application Clinical Correlations with Positive DAT

Antibody Detection 1. Hemolytic Disease of the Newborn (HDN)
Crossmatching Occurs in Rh (-) mother with Rh (+) fetus
Antibody screen 2. Hemolytic Transfusion Reaction (HTR)
Antibody panel Recipient’s antibody coating Donor’s RBCs
Antibody Titration 3. Autoimmune Hemolytic Anemia (AIHA)

Reaction Medium and Incubation Time

ALBUMIN LISS PEG

Medium (22% Bovine Serum Albumin) (Low Ionic Strength Solution) (Polyethylene glycol)
NSS

Incubation 30 MINS 10-15 MINS 10-15 MINS 30-120 MINS

 Blood Donation
Donor Screening
PHYSICAL MEDICAL HISTORY
REGISTRATION
EXAMINATION QUESTIONNAIRE

TEMPORARY - unable to donate for a

limited period of time

DONOR INDEFINITE - unable to donate for an

DEFERRAL unspecified period of time

PERMANENT - will never be eligible to

donate blood for someone else.
AUTOLOGOUS DONATION
Donor who donates blood for his or her use
Donor-Patient

TYPES OF AUTOLOGOUS DONATION

Preoperative Collection/ Predeposit donations
Occurs during 5-6 weeks immediately preceding a scheduled elective surgical procedure
Immediate Preoperative Hemodilution
Performed immediately prior to beginning the surgical procedure
Intraoperative Collection
Involves collecting shed blood from surgical site
Postoperative Collection
Collected from a drainage tube placed at the surgical site

APHERESIS DONATION
uses an instrument/machine to selectively collect and separate a specific
component
Plateletpheresis
Platelets obtained by an apheresis procedure provide the equivalent of 6-8 whole-blood-derived
platelets
Plasmapheresis
Each unit is the volume equivalent of at least 2 whole-blood-derived plasma units
Leukapheresis
Typical therapeutic dose is at least 1 x 10^11 granulocytes each day for 5 consecutive days
In order to collect a large enough volume of leukocytes:
Hydroxyethyl Starch (HES)
Corticosteroids
Granulocyte Colony stimulating Factor (GCSF)
DONOR REACTIONS

COLLECTION FREQUENCY
2 RBC
Plasma (Frequent)
Plasma (Infrequent)
16 weeks
Every 2 days (no more than 2x in 7 days)
Every 4 weeks (no more than 13x/year)
1. MILD REACTIONS
fainting, nausea or vomiting,
hyperventilation, twitching, muscle spasm
2. MODERATE REACTIONS
Mild reactions in addition to loss of
consciousness

Every 2 days (no more than 2x in 7days; no

3. SEVERE REACTIONS
Platelets (single unit) Convulsions
more than 24x in 1 year)

Platelets (double or triple units) Every 7 days 4. HEMATOMA

Granulocytes Every 2 days Patients are asked to keep pressure on
punctured site for 5-10 minutes
 Pre-Transfusion testing
GOALS
Ensure blood components transfused are compatible with the recipient’s blood
Ensures maximum red cell survival
Prevent disease transmission

REQUEST IN BLOOD BANK

Hold Clot
Clotted Sample held in transfusion service but is not tested at all
Type and Hold
ABO and Rh typing done but no other testing
Type and Screen
ABO, Rh Typing done, antibody detection performed
Type and Crossmatch
Same as type and screen but adds crossmatch
Maximum Surgical Blood Ordering Schedule (MSBOS)
Generally is a list of surgical and other procedures
 Procedures in Pre-transfusion Testing
Patient Specimen
Requisition Form
Identification Collection

Component Collection of Donor Sample

Selection Sample Evaluation

TYPE AND THE

SCREEN CROSSMATCH

DONOR CLASSIFICATION

Autologous blood
patient donates blood for his/her own surgery
Directed donation
blood collected from individuals designated by the patient as acceptable donors
Allogeneic blood
comes from the general population
Causes of Incompatible Crossmatches

Antibody Screen Negative

Antibody Screen Positive
Antibody Screen Positive, Autocontrol Positive

SPECIAL CIRCUMSTANCES

Emergency Transfusion
Physician will need to sign an emergency release form, waiving compatibility testing
Massive Transfusion
Total blood volume has been replaced with donor blood within 24 hrs
Intrauterine Transfusion
Compatibility testing performed using mother’s serum sample
Neonatal Transfusions (Infants younger than 4 months)
Donor unit: compatible with mother and baby.
Autologous Transfusion
removal and storage of blood or components from a donor’s possible use at a later time
 REFERENCES
Video Discussions posted in Google Classroom by Ms. Sarah Jane V. Denia, Mr. Rhoyette Cuyo,
and Cristine Joy Versoza
FINAL PROJECT IN IMMUNOHEMATOLOGY

Submitted by:

Maricel D. Lorque
BSMT 3-2 Cluster 1