IMMUNO-SERO

MIDTERM REVIEWER
Other Major Blood Groups: Part 1

LEWIS SYSTEM
LEWIS (007) SYSTEM

- Lewis gene (Le) codes for the production of

fucosyltransferase enzyme. (makikita sa
membrane ng RBC)
- Erythrocytes acquire the lewis phenotype by
adsorbing Lewis substances from the
plasma. Rather than being membrane-
bound antigens. (Galing plasma pero
madedevelop sa erythrocyte)

TYPE 1 PRECURSOR CHAIN

LEWIS ANTIGEN

- Cord blood and red cells from newborn

- L- fucose: dominant sugar infants phenotype as Le(a-b-)- ito yung
subgroup ni lewis
- Attached in Beta 1-3 of galactose and N-Acetyl
- Decrease in expression of Lewis antigen has
glucosamine
been demonstrated on red cells from many
pregnant women.

JD 💉
 Ss ANTIGENS

- Located on Glycophorin B
- Also known as Ss-sialoglycoprotein (SGP)
- Amino acid at position 29 on GPB is critical to
antigen expression:
PHENOTYPES AND FREQUENCIES IN THE LEWIS - S has methionine,
SYSTEM - s has threonine.
- Well developed at birth.
- Less easily degraded by enzymes.

LEWIS ANTIBODIES

- Considered naturally occuring.

- IgM in nature.
- Activate the complement and can cause in
vivo and in vitro hemolysis. (in vivo ay sa
loob na pwedeng mag cause ng hemolytic
disease of newborn while in vitro ay
magkakaroon ng hemolyzed sample after
extraction since sa labas ito)

MNSs (002) BLOOD GROUP

SYSTEM
MN ANTIGEN

- Found on glycophorin A of cell membrane.

- Also known as MN-sialoglycoprotein.
- Differ in their amino acid residue at positions
1 and 5.
- M has serine and glycine
- N has leucine and glutamic acid
- Easily destroyed or remove by enzymes.
- Well develop at birth
o Important in paternity studies of
infants and young children ANTIBODY
Anti-M

- Naturally occurring cold reactive IgM or IgG.

- pH dependent: React best at pH 6.5 (acidic)
- Usually do not bind complement.
- Other examples react only with red cells
exposed to glucose solutions.
- Rarely causes hemolytic transfusion
reactions or HDN.

ANTI-M LECTIN

- Iberis amara
- Iberis umbellate

JD 💉
 - Iberis semperivens
- Iberis aurantiaca

Anti-N

- Naturally occurring cold reactive IgM or IgG.

- Rare cause of HDN
- Anti-N like antibody found in renal patient
dialyzed with formaldehyde sterilized
equipment.

ANTI-N LECTIN
FREQUENCY OF P SYSTEM PHENOTYPES
- Bauhinia variegata
- Bauhinia bonatiana
- Bauhinia purpura
- Bauhinia candicans
- Vicia graminea

Anti-S and anti-s

- Most examples are IgG, reactive at 37°C and

the antiglobulin test phase.
- Implicated with severe hemolytic transfusion
reaction with hemoglobinuria, and HDN

ANTIHUMAN GLOBULIN TEST

ANTIGEN
P1 ANTIGEN

- Found on fetal rbc as early as 12 weeks,

weakens with gestational age.
- Deteriorates rapidly on storage.
- P1-like antigen found in plasma, and
droppings of pigeons and turtledoves, egg
white of turtledoves.
- P1 substance
o Hydatid fluid,
o Extracts of Lumbricoides terrestris,
Ascaris suum.

Direct globulin test: in vivo antibody

Indirect globulin test: in vitro antibody ANTIBODY

ANTI-P1

- Common, naturally occurring IgM antibody

in the sera of P2 individuals.
- Cold-reactive saline agglutinin.
- Strong anti-P1 was observed in individuals
infected with HYDATID DISEASE.
- Associated with fascioliasis, Clonorchis
sinensis, and Opisthorchis viverrini
infections.

Anti-PP1Pk

P System Phenotypes, - Originally called anti-Tja

- Predominantly IgM, sometimes IgG.
Antigens, and Antibodies - Reacts over a wide thermal range.
- Associated with spontaneous abortions in
P SYSTEM PHENOTYPES, ANTIGENS, AND
early pregnancy.
ANTIBODIES
Anti-P

- Naturally occurring alloantibody in the sera

of all Pk individuals.
- Alloanti-P is rarely seen in the blood bank.
o Very significant because it is
hemolytic with a wide thermal
range of reactivity.

JD 💉
 - Anti-P specificity is found as an IgG
autoantibody in patients with Paroxysmal
Cold Hemoglobinuria
- Antibody activity is biphasic – it attaches to
red cells in the cold and lyses them as they
warm.
- Demonstrated by the Donath-Lendsteiner
test.
ANTI-P (BIPHASIC)
ANTI-PK
- Isolated from some examples of anti-PP1Pk
by selective adsorption with P1 cells.
- Has been reported in the serum of P1
individuals with biliary cirrhosis and
autoimmune hemolytic anemia.
I (207) BLOOD GROUP
COLLECTION
Ii ANTIGENS
- At birth, infant red cells are rich in i; I is almost
undetectable.
- During the first 18 months of life, the quantity of i
slowly decrease as I increase.
- Adult red cells are rich in I and have only trace
amount of i antigen.
Increase i Antigen = HEMPAS
- Hereditary Erythroblastic Multinuclearity with
Positive Acidified Serum Test
Rare I Adult or I Negative Phenotype
- Individuals who do not change their i status after
birth.
ANTIBODY (Anti-I)
Anti-I
- Common autoantibody that can be benign or
pathologic.
- Demonstrates strong reactions with adult cells and
weak reactions with cord cells.
- Not associated with HDN because the antigen is
poorly expressed on infant red cells.
Benign Anti-I
- Found in the serum of many normal healthy
individuals.
- Not associated with in vivo red cell destruction.
- Weak, naturally occurring, saline-reactive IgM
agglutinin.
- Usually reacts only at 4°C
Pathologic Anti-I
- Potent IgM agglutinins with higher titers and
broader thermal range of activity, reacting up to 30
to 32°C.
JD 💉
- Attach in vivo and cause autoagglutination and
vascular occlusion (Raynaud’s phenomenon) or
Antibodies to k,Kpb, Jsb, and Other High-Frequency
intravascular hemolysis.
Kell antigens
- Associated with CAD, CAS and PAP. (Primary atypical
- Antibodies to high-frequency Kell antigens are rare,
pneumonia {PAP}, Cold autoimmune disease {CAD})
because so few people lack the antigen.
Autoanti-I
McLeod Phenotype
- Production of autoanti-I may be stimulated by
- Rare phenotype with decreased Kell system antigen
microorganisms carrying I-like antigen on their
expression and abnormal red cell morphology.
surface.
- Associated with Chronic Granulomatous Disease.
❖ Mycoplasma pneumoniae
FREQUENCIES OF COMON KELL PHENOTYPE
❖ Listeria monocytogenes

Autoanti-i

- Most autoanti-i are IgM.

- Reacts best with saline-suspended cells at 4°C.

- Potent examples are associated with:

✓ Infectious mononucleosis
✓ Disease of reticuloendothelial system
o Alcoholic cirrhosis, myeloid
leukemia, reticulosis

- IgG anti-i has also been described and has been

associated with HDN.

DUFFY (008) BLOOD GROUP

SYSTEM
KELL (006) BLOOD GROUP
SYSTEM Fya and Fyb Antigens

- Identified on fetal red cells as early as 6 weeks

Kell Antigens
gestational age and are well developed at birth.
- Immunogenic, K is rated second only to D in terms
- Destroyed by common proteolytic enzymes.
of immunogenicity. (Immunogenic- kailangan
maexpose muna sa antigen para ma develop ang Anti-Fya and Anti-Fyb Antibodies
characteristics)
- Usually IgG and react best at the antiglobulin
- Well-develop at birth. phase.
- Expression very weak on McLeod phenotype cells. - Activity is enhanced in a low ionic strength
medium. (ginagamitan ng low ionic strength saline
Anti - K
LISS sa lab)
- Common antibody encountered in blood bank.
- Do not react with ENZYME-TREATED red cells.
- Usually an IgG antibody reactive in the antiglobulin
- Associated with hemolytic transfusion reactions,
phase.
although hemolysis is not often severe.
- Made in response to antigen exposure through
FREQUENCY OF DUFFY PHENOTYPES
pregnancy and transfusion.

- Implicated in severe hemolytic transfusion reactions.

- Associated with severe hemolytic disease of the

newborn.

Antibodies to Kpa, Jsa, and Other Low-Frequency Kell

antigens

- Antibodies are rare because so few people are

exposed to the antigen.

- Most often detected through unexpected

incompatible crossmatches or cases of HDN.

JD 💉
 - May be IgA, IgM, or IgG.

- Most are IgG (often IgG4) although IgM and IgA

antibodies have been noted.
KIDD (009) BLOOD GROUP - Reactive at 37°C and the antiglobulin phase.
SYSTEM
- Made in response to pregnancy or transfusion.
a b
Jk and Jk Antigens
- Implicated with shortened survival of transfused
- Detected on fetal red cells as early as 11 weeks for cells and posttransfusion jaundice.
Jka and 7 weeks for Jkb .

- Well develop at birth.

Minor Blood Groups
- Not altered by enzymes.

Frequencies of Kidd Phenotypes

Diego Chido/Rodgers
Cartwright Gerbich
Xg Cromer
Scianna Knops
Dombrock Indian
Colton

ANTIBODY CHARACTERISTICS IN A NUTSHELL

Anti - Jka and Anti - Jkb Antibody

- Have notorious reputation in the bloodbank.

- Immune antibodies, made in response to pregnancy

or transfusion.

- Detected in the antiglobulin phase.

- Common cause of hemolytic transfusion reactions,

especially of the delayed type.

Diego blood group system (010)

- Dia antigen has served as a useful tool in
anthropologic studies of Mongolian ancestry.

CARTwright blood group system

(011)
- YT antigens have been located on erythrocyte
LUTHERAN (005) BLOOD acetylcholinesterase, which is an enzyme involved in
neurotransmission.
GROUP SYSTEM
Lua and Lub Antigens

- Poorly developed at birth and do not reach adult XG blood group system (012)
levels until age 15.
- Gene that encodes for the Xg allele is located on the
Anti - Lua Antibodies short arm of the X chromosome (sex chromosome
pair number 23)
- Most are naturally occuring saline agglutinins that
react better at room temperatures than 37°C. - Difference in the frequency of the Xga antigen is
noted between the sexes:
- Few react at 37°C by indirect antiglobulin test.
✓ 89% female

JD 💉
 ✓ 66% female

CROMER BLOOD GROUP

SCIANNA BLOOD GROUP SYSTEM (021)
SYSTEM (013) - Antigens are carried by decay accelerating factor
(DAF), which is involved with the regulation of
- SC system is composed of the Sc1, Sc2, and Sc3
complement activation by accelerating the decay of
antigens.
the C3 and C5 convertase.
- Anti-Sc3 is characterized as IgG, red cell-stimulated,
and reacting in the IAT phase of testing.

o Causing mild transfusion reaction

KNOPS BLOOD GROUP
SYSTEM (022)
DOMBROCK BLOOD GROUP - Alleles for the KN blood group have been located on
chromosome 1 (Rh), with the antigens residing on
SYSTEM (014) complement receptor one (CR1).

- Doa, Dob

- Gregory (Gya), Holly (Hy), and Joseph (Joa) INDIAN BLOOD GROUP
- Anti-Doa, Anti-Dob are described as IgG, red cell- SYSTEM (023)
stimulated antibodies, react primarily in IAT with PEG
or enzyme enhancement. - IN antigens are carried on the hematopoietic
isoform of the CD44 marker, which is known for its
- Anti-Gya, Anti-Hy, and Anti-Joa are IgG, red cell immune adhesion properties.
stimulated antibodies and IAT reactive.

HUMAN LEUKOCYTE ANTIGENS (HLA) DETECTABLE

COLTON BLOOD GROUP ON RBCs

SYSTEM (015) - Bga antigen, corresponds with HLA-B7, Bgb with HLA-
B17, and Bgc with HLA-A28
- CO antigens have been located on the transport
protein known as channel-forming integral protein High Incidence Antigens
(CHIP), which forms the primary erythrocyte water
- Antigens of high frequency are defined as occuring
channel and is responsible for water permeability.
in 99.9% of the population.

- Examples of high-frequency antigens that are

CHIDO/rodgers Blood group
system (017)
- CH/RG antigens were associated with the human
leukocyte antigen (HLA) system. (concentrating on
the major histocompatibility complex, pagdating sa
organ donation, pwedeng magkaroon ng reaction)
- Alleles for RG and CH have been located on two
closely linked genes known as C4A and C4B on
chromosome 6.
- Anti-Ch/Rg antibodies were collectively grouped as
HIGH-TITER, LOW-AVIDITY (HTLA), along with other
antibodies sharing common serologic properties.
apparently unrelated to principal blood group
systems include:
o Augustine (Ata), Cromer (Cra), Ena, Gerbich
(Ge), Gregory (Gya) and Holley (Hy), Jacobs
(Jra), Joseph (Joa), Langereis (Lan), Oka, and
Vel (Ve)
Low-Incidence Antigens
- Low-incidence antigens unrelated to the principal
blood group systems are defined as antigens with an
incidence of less than 1%.
- Examples of this type of antigen include the Wright
blood group system, which is independent of all other
blood group systems, as well as Swann (Swa), By, Mta,
and Tra antigens.

HIGH-TITER, LOW-LOW AVIDITY (HTLA) Antibodies

GERBICH BLOOD GROUP
- Antibodies exhibit reactivity at high dilutions of
SYSTEM (020) serum, but the strength of agglutination is weak at
any dilution.
- GE antigens are inherited on chromosome 2 and are
expressed on glycophorins C (GPC) and/or D (GPD)

JD 💉
- Antibodies formerly classified as HTLAs owing to
similar serologic reactivity:

o Anti-Ch (Chido)
o Anti-Rg (Rodgers)
o Anti-Kn (Knops)
o Anti-JMH (John Milton Hagen)
o Anti-Yka (York)
o Anti-Csa (Cost)
o Anti-McC (McCoy)

Detection and Identification of Antibodies

a. Enzyme techniques
b. Elution
c. Adsorption
d. Neutralization
e. Chemicals

Enzyme techniques

Enhanced:

o Rh, Le, I, Kidd, P, Anti-P1

Inactivated:

o MNS, Duffy, Xg, Ch, Rd, Yt

Antibody Screening, WHY?

Indications: For detection of irregular antibodies

Antibody Screening (non-ABO)

• Pre-transfusion tests : before compatibility

Compatibility Testing Protocols
ABO GROUPING
- Most critical pretransfusion serologic test.
- If the patient’s ABO group cannot be
satisfactorily determined and immediate
transfusion is essential, group O packed
red cells should be utilized.
Rh TYPING
- If Rh type of the recipient cannot be
determined and transfusion is essential, Rh-
negative blood should be given.
ANTIBODY SCREENING
- Purpose: To detect as many “clinically
significant antibodies” (outside of ABO
antibodies) as possible.
- “Clinically significant Abs” – refers to Abs
that are reactive at 37ºC and/or in the AHG
test and are known to have caused a
transfusion reaction or unacceptably short
survival of the transfused red cells.
testing, kailangan malaman kung may
antibody present sa patient na hindi
kailangan at meron sa donor. Kung + si donor
at patient sa antibody na yon, magkakaroon
ng hemolysis. Needed na neg yung blood
donor.
• Antenatal screening: HDN, bago mag give
birth si mother
• Donor units: examine the donor units for the
type of antibody para malaman kung
ittransfuse ba ang blood.
• HDN
• On request,e.g. Immune complex test (ICT)
for Autoimmune hemolytic anemia (AIHA)
Antibodies regarded as always being potentially
clinically significant:
ABO Duffy
Rh Kidd
Kell SsU
- Antibody screening is done by testing the
patient/donor serum against screening cells, a panel
of commonly encountered and clinically significant
antigens. Screening cells are Group O that have
known antigens present. Commercially prepared
available sets of screening cells contain D, C, E, c, e,
M, N, S, s, Lea, Leb, P K, k, Fya, Fyb, Jka, and Jkb
antigens. Testing is performed in three consecutive
phases using patient serum:

o Immediate Spin in saline at Room Temp.

(kapag may nag (+) automatic ay IgM-

JD 💉
 reactive antibodies na malamig known as
cold reactive antibodies)
o 37ºC incubation with enhancement medium
(e.g. albumin, LISS, PEG) (sa common setting
ang ginagamit ay LISS) (22% bovine albumin-
colorless)
o AHG Phase. (color green ang anti sera)

- double dose: Ss/MN/Kk kapag both present. Kapag

isa lang ay single dose.

ANTIBODY SCREENING PROCEDURE

Auto control: Patient serum – Patient red cells (testing
for autoimmune if may autoantibodies)

Kapag negative sa abtibody screening, proceed na sa

cross matching

Sa patient 2 may positive reaction (agglutination)

meaning may presence ito ng alloantibodies. Kapag
positive, screen cell na umaabot ng 11.

A+ si patient and may positive reaction sa phase 2,

meaning mag alloantibodies which is IgG.

• AHG
Antibody Identification
• Enzyme
• --------------------- - If the Antibody Screen is reactive, the antibody
• One-cell pool (donors) specificity must be determined.
• Two cells
- So safe blood can be administered to the Recipient.
• Three cells (recommended)
• Six cells - 11 reagent panel cell are to be used for
identification.
Enzyme treatment: checking

Kapag + sa coombs test dapat (-) sa enzyme kasi may

neutralization na nangyayari. If (-) naman sa coombs
test, dapat maging (+) sa enzyme kasi ineenhance ni
enzyme yung antibody na present.

Coombs test: AHG phase

Antibody Screening

Antibody Screening and Identification

Techniques to enhance antigen-antibody reactions

thus facilitating antibody identification include

b.1 enzyme treatment with ficin, papain,

trypsin, or bromelain.

JD 💉
Enhanced: Kidd, Rh, Ii, P, Lewis Question: Ano ang present sa patient sa sample 1 ?

Destroyed: MNS, Duffy Answer: Meron siyang allo antibody na IgM

b.2 Increasing the amount of

serum to increase the number of available antibody
Question: Ano ang present sa patient sa sample 2 ?
molecules
Answer: Meron siyang alloantibody na IgG
b.3 Lengthening incubation time

Ficin: 2 portions

Question: Ano ang present sa patient sa sample 3 ?

Answer: Meron siyang alloantibody at autoantibody

which could be considered IgG warm autoimmune
hemolytic anemia (WAIHA)

Question: Ano ang present sa patient sa sample 4 ?

Papain: 3 portions Answer: Meron siyang IgM cold autoimmune

hemolytic anemia (CAIHA)
- Ang purpose ay para madetached and
magkaroon ng negative reaction.

Antibody Identification

Una check kung IgM or IgG based sa temperature,

based sa table may 2+ reaction sa RT so ito ay IgM.
Iccheck kung anong pattern correspond to AHG na IgG: S,s,Jka, Jkb, Fya,Fyb, K, D,C,E,c,e
nakalagay sa gilid. In this case, Xg and M positive si
patient. IgM: M,N,P1,Lea, Leb, I

Based sa table, si patient ay merong M at P1. Kaso

dalawa ang present so need pa ng neutralization (para
madissolve yung isa sa kanila)

Sa babang table naman, positive kay c so need

magprepare ng blood bag na negative kay c.

- Kapag positive sa IS (room temp) ay IgM

- Kapag positive sa 37’C at AHG ay IgG
D ang antibody

JD 💉
LISS/Coombs phase (ID-DiaPanel) - ELUTION : used to dissociate IgG Abs from
sensitized red cells

: the recovered antibody, eluate, can be

tested like serum to determine the antibody’s
specificity

: techniques include heat, freeze-thaw

process, use of organic solvent, acid eluates, or by
Enzyme phase (ID-DiaPanel-P) using ZZAP or chloroquine diphosphate

*ZZAP- mixture of DTT and papain that is used to

remove Ab from sensitized red cells and to enzyme
treat them at the same time

*Chloroquine diphosphate- reagent used to remove

IgG Abs from the surface of sensitized cells;
inactivates Bg antigens

Interpretation and Remarks

Antibody Screening

o Probable ab : Anti-D, k, Kpb, Jsb, P1, M, N, s,

Lub

ID : Anti-M

Antigen typing : M-N+

MNS antibodies show weaker ( or NEGATIVE) reaction

ELUTION Test / Principle
in enzyme than in Coombs test

Antibody Screening and Identification

Other techniques may be used to eliminate clinically

insignificant reactions and make identification of
significant antibodies easier.

c.1 Use of AET, DTT, and ZZAP

which inactivates some antigens especially Kell.

c.2 Prewarm procedure. Clinically

insignificant cold antibodies may be removed by this
technique. Patient serum, reagent red cells and
enhancement medium can be warmed separately at
37°C for 5-10 minutes prior to mixing

c.3 Use of sulfhydryl or thiol

reagents (DTT and 2-ME) which denature IgM
antibodies by breaking disulfide bonds.

c.4 Use of adsorption and elution

techniques to remove unwanted antibodies such as
cold or warm autoantibodies, or to help resolve c.5 Another technique for facilitating antibody
multiple antibodies identification is NEUTRALIZATION.

Commercial substances are available to

neutralize or to inhibit reactivity of some antibodies.
ADSORPTION & ELUTION TECHNIQUES
Sources of Substances for Neutralization of
- ADSORPTION: used to remove unwanted antibodies Antibodies:
from SERUM.
Hydatid cyst fluid – anti-P1
- If an autoantibody such as I, H, or IH are defined, it
can be adsorbed onto the patient’s enzyme Plasma or serum w/ Le substances – anti-Lea & anti-
pretreated cells at 4ºC. Rabbit cells may also be used Leb
as adsorbents for anti-I since they are rich in I antigen. Pooled serum or plasma – anti-Chido, anti-Rogers

Urine – anti-Sda

JD 💉
Saliva of “secretors” – anti-ABH Ratio: 2 drops of serum and 1 drop of red cell
suspension
Human milk – anti-I
Purpose:

1. Final check of ABO compatibility between patient

and donor to prevent transfusion reaction.
Compatibility Testing 2. Detects presence of antibody in patient’s serum
that will react to donor’s RBC that is not detected n
antibody screen.
- Crossmatching in other term
3 Phases of Crossmatching
COLLECTION AND PREPARATION OF SAMPLES
1. Immediate Spin in saline at RT - Detects IgM
1. Patient Identification
2. Thermophase/37ºC incubation for 30 minutes with
- name, gender, occupation, date ng surgery enhancement medium (e.g. albumin, LISS, PEG) –
Detects IgG
2. Collection. SERUM: preferred specimen for
compatibility testing. Hemolysis should be avoided. 3. AHG Phase after washing incubated cells with
saline.
Why SERUM and NOT PLASMA?
- Antihuman globulin: antisera that is made up from a
- Plasma may cause small fibrin clots to form which
human serum then injected to rabbit tapos
may be difficult to distinguish from true agglutination.
magkakaroon ng neutralization process wherein mag
(magkakaroon ng false positive agglutination:
s-sensitized yung kanyang serum then pwede nang
nagkaroon ng agglutination pero hindi dapat)
kuhanin yung blood
- Plasma may inactivate complement so that
antibodies may not be detected. (complement
pathway ay para sa cell lysis. Paano nagkakaroon ng
cell lysis? By antigen-antibody reaction with the help
of classical pathway. Kung inactivated ang
complement, walang makakadetect na merong
antibody sa serum sample)

3. Age of Specimen. The freshest sample possible

should be used for compatibility testing. Specimens
must be less than 3 days old if the patient has been
transfused or pregnant within the past 3 months. (the
sample could be stored until 7 days with refrigerated
temp. However, as much as possible mas Maganda A & B: Incompatible (agglutination)
kung fresh, meaning mas preferred yung less than 3
days) C & D: Compatible (No agglutination)

4. Sample Storage. The AABB requires that patient Check cells/Coombs control cells
samples must be stored between 1-6ºC (refrigerated - (IgG sensitized cells) should be added to tubes that
temp) for at least 7 days after transfusion. demonstrate no agglutination.

- For results to be considered valid, agglutination

Crossmatching must occur.

METHODS AHG PHASE OF CROSSMATCHING

Slide- emergency cases

Tube- broad spectrum compatibility test (lab)

(preferred)

Gel method

-------------------------

(MJ) MAJOR X-MATCH: Donor’s cells + Recipient’s

serum
- yung AHG reagent ay may dalawang type (Poly and
(MN) MINOR X-MATCH: Donor’s serum + Recipient’s
Mono) in this case poly specific yung ginamit. Kapag
cells
may AHG na sa specimen at iccentri for 1 minute, then
(AC) AUTO-CONTROL: Patient serum + Patient red cell check for agglutination. Kapag may agglutination,
there are still an incomplete antibodies that is not
bound to each other kaya bin-bound na ng AHG.

JD 💉
 2. PERFLUOROCHEMICALS (PFCs)

- (examples: Fluosol-DA-20 , Oxygent)

- excellent gas (O2 and CO2) solvents

Dahil dito, kailangang magkaroon ng positive or
agglutination reaction doon sa nag negative kanina
para ma consider na valid.

Donor Selection, Blood Collection &

Component Preparation

DONOR SELECTION, BLOOD COLLECTION &

COMPONENT PREPARATION

DONOR SELECTION, BLOOD COLLECTION &

COMPONENT PREPARATION

1. PERSONAL INFORMATION
Reporting of Results
a) Donor’s name
A compatible crossmatch is indicated by absence of b) Donor’s address, phone no.
agglutination and/or hemolysis at any stage of the c) sex
crossmatch. The absence of agglutination indicates d) age (17 years old and above, 17 must with
that the patient has no demonstrable antibodies with parent’s consent)
a specificity for any antigen on donor’s RBC. e) date of donation
f) donor’s consent
Troubleshooting Incompatible Crossmatches
g) donor’s occupation
h) race
i) time of last meal

2. PHYSICAL EXAMINATION

a) General Appearance. (to be examined by

physician)
b) Weight. 110 lbs (50 kg)
If donor is less than 110 lbs:
Amount of blood to be drawn:
The Future of Compatibility Testing

- Red cell/Blood substitutes***

- Biochemical modification of non-O blood

- Galvanic biosensor – energy measured

- Dipstick method of typing (ex. Eldoncard Blood

typing kit)

- Dry plate method c) Temperature: Orally should not exceed

BLOOD SUBSTITUTES
- substances that are able to carry oxygen in the
absence of intact red cells
1. Stroma-free Hb solns/Hemoglobin-Based Oxygen
Carriers (HBOCs) (e.g. PHP, PEG-Hb, Hemolink,
Polyheme, HemAssist, Hemopure, Optro)
99.5ºF or 37.5ºC
d) Pulse: 50 to 100 beats per minute
e) Blood Pressure: Systolic no greater than
180mmHg, diastolic no greater than 100
mmHg (Hypertensive phase 1: 140/100)
f) Hematocrit and Hemoglobin: 38% Hct (12.5
g/dL Hb)

JD 💉
COPPER SULFATE METHOD (CuSO4) - (+) serologic test for syphilis

Principle: A drop of whole blood when dropped in a - donors who have traveled to areas considered
solution of CuSO4 , endemic for malaria

which has a given specific gravity, will maintain its (don’t defer a donor who started antimalarial therapy
density for approximately 15 seconds. in preparation for travel to areas endemic for malaria)

Specific gravity of CuSO4 is 1.053 which is equivalent - rabies vaccine

to 12.5 g/dL
- sexual contact with any person who has high risk of
(gagamitan ng beaker containing 30mL copper sulfate exposure to HIV
reagent na pwede lang sa 25 donors) (kapag nagsink
Two months (8 weeks/ 56 days)
yung blood ay acceptable)
- whole blood donation

6 Weeks
3. MEDICAL HISTORY
- pregnant: deferred during normal pregnancy and 6
DONOR DEFERMENT:
weeks following a third trimester delivery (kapag CS
Indefinitely or permanently ay 1 year)

a) 1. history of viral hepatitis - 1st and 2nd trimester abortion or miscarriage need
- (+) HBsAg not to be a cause for deferral (after 2 moths kapag
- reactive for Anti-HBc nagkaroon ng abortion)
- past/present evidence of Hepatitis C
One Month (4-week deferral)
infection
- donor involved in post transfusion hepatitis - donors taking Accutane (isotetinoin for acne therapy
b) history of jaundice of unknown cause is also a potent teratogen)
c) past/present abuse of self-injected drugs
d) cancer - Vaccination: Rubella (German measles) Varicella
e) abnormal bleeding tendencies zoster (chicken pox)
f) cardiopulmonary diseases
Two-week deferral
g) leukemia, lymphoma
h) high risk sexual behavior - Vaccination: attenuated virus vaccines
i) high risk occupation (e.g. prostitute)
j) Chaga’s disease a. Mumps e. Yellow fever
k) Babesiosis b. oral polio (Sabin) f. Influenza (live virus)
l) those receiving growth hormone
(Creutzfeldt-Jakob dse.) c. Rubeola (measles)
m) symptoms of AIDS Related Complex (ARC),
d. Smallpox)
HIV/AIDS
n) donor’s taking Tegison for psoriasis because 3 Days
its potentially teratogenic
o) Active pulmonary TB - tooth extraction or dental work (mas safe kung after
a week)

48-hours
Three Years Deferral
- donors participating in pheresis program (pheresis
- those infected with malaria ay when the RBC is separated into platelet and plasma
and ito ay mapupunta sa blood bag while yung RBC ay
- visitors, immigrants or refugees from an area
babalik sa patient)
considered endemic
Until signs and symptoms are gone
for malaria/ residents of area endemic for malaria
- donors who have active cold or flu symptoms must
12-month deferral
be deferred until the symptoms are gone
- close contact to patient with hepatitis
No deferral
- donors who received blood or blood products, an
-donors who had vaccinations of the following killed
organ or tissue transplant
viral, bacterial, rickettsial vaccines or toxoids
- tattoo, ear and skin piercing
diphtheria tetanus
- those who have received HBIg bec, it is given for
cholera Hepa B vaccine
exposure to possible infection and it may delay the
onset of symptoms of disease pertussis typhoid

- donors who have had or been treated for syphilis or RMSF influenza (killed)
gonorrhoea

JD 💉
injectable polio vaccine (Salk)

Condition A Deferral

Condition F Deferral

Condition B Deferral

Condition G Deferral

Condition c Deferral

Condition H Deferral

Condition D Deferral

JD 💉
Condition I Deferral Condition R Deferral

Condition M Deferral Condition S Deferral

Condition T Deferral

Condition O Deferral

Condition U Deferral

Condition P Deferral

JD 💉
ACCEPTABLE DRUGS:
*Allergy shots
*Analgesics (pain medications such as aspirin)
*Anorexics (diet pills)
*Antibiotics (Completed medication and is symptom
free) (after week of taking antibiotics)
*Antidepressants
*Antacids
*Antihistamines
*Antihypertensive agents
*Antispasmodics
*Birth control pills
*Corticosteroids
*Diuretics
*Eye medication
*Fertility agents (if not pregnant)
*Gout medication (must be symptom free)
*Hormones
*Muscle relaxants
*Sedatives
*Thyroid
*Tranquilizers
*Vitamins
Medication Guidelines
- Acetaminophen (e.g. Tylenol): may be taken in
normal moderate doses before any Blood donation
- Accutane: four-week deferral
- Allergy medication: can donate
- Antibiotics: 72-hour deferral after infection is healed
- Anti-inflammatory drugs (Advil, Ibuprofen, Motrin
and Naprosyn): may not be taken within 24 hours
before a platelet donation (some other rules may
apply)
- Aspirin-containing products or Feldene and Lodine
XL: may not donate within 36 hours before platelet
donation
- Birth control pills: can donate
- Blood pressure medication: can donate under
present FDA and American Red Cross standards in
force
- Depression medication: can donate
- Diabetic medication - Injected bovine (beef) insulin
since 1980; can not donate
- Diet pills: can donate
- Diuretics: can donate
- Female hormone pills: can donate
- Any human pituitary-derived hormone (i.e. growth
hormone): can not donate
- Soriatane (Acitretin): three-year deferral
- Tegison (used to treat a severe skin disorder): can
not donate if ever taken
- Thyroid medication: can donate if stabilized
QUESTION!!!
According to AABB standards, which of the following
donors may be accepted as a blood donor?
a. Hip replacement 5 months ago (general
surgery)
b. Spontaneous abortion at 2 mos. pregnancy,
3 mos. Ago (magkasunod na abortion)
c. Resides with a known hepatitis patient
(incubation period pa)
d. Received a blood transfusion 22 weeks
previously (naka 5 months na)
HEMAPHERESIS DONOR SELECTION
Hemapheresis – type of blood donation where whole
blood is withdrawn either from a donor or patient
wherein after removal, separation, and retention of
the desired cellular elements or plasma, the
remaining products are recombined and returned
to the donor or patient.
Common: plasma and platelet
ADDITIONAL DONOR GUIDELINES
Other term for voluntary donor is allogenic donation.
Unlike voluntary donor, we need to wait 8 weeks to
donate.
1. At least 48 hours is the elapsed time after
hemapheresis donation.
2. A donor must not exceed more than two times in
a week or 24 times in a year unless otherwise allowed
by bloodbank physician. (the donor should know when
is the last donation)
3. A donor must be tested to detect cytopenia. (
occurs when one or more of your blood cell types is
lower than it should be.)
4. If a donor donates whole blood, at least 8 weeks
must be elapsed before he can donate for pheresis.
5. Extracorporeal blood must not exceed 15% of the
donor’s total blood volume.
6. If platelet pheresis is to be performed a donor must
have above 150 x 109/L platelet count. (normal value
of platelet: 150,000 to 400, 000)
7. Possible adverse reactions to HES, (Hydroxyethyl
starch (HES) is the most common red blood cell
sedimentation agent used to purify neutrophil
collections for granulocyte transfusions) steroids
and/or heparin must be determined. These
substances are used in the apheresis procedures.
AUTOLOGOUS DONOR SELECTION
Autologous Donor – one who is donating blood for his
or her OWN future use.
Autologous blood – the safest blood possible for
transfusion
a. no risk of disease transmission;
alloimmunization to red cells, platelets,
wbcs, or plasma proteins; transfusion reactions
b. phlebotomy process stimulates the BM to increase
cell production
c. decreases the need for allogeneic blood and may
actually increase the supply for allogeneic blood
supply.
CRITERIA
No age limit. (allogenic: 18-65 years old)
No strict weight requirements (allogenic:
110 pounds ang acceptable)
Hemoglobin/hematocrit – should not be less
than 11 g.dl and 34% (allogenic: 12.5g/dL and 38%
ang hematocrit) the patient needs to be transfused if
the hemoglobin 7 g/dL.
JD 💉
 Frequency – Donations should not be more
frequent than every 3 days and the final donation
must be completed at least 3 days prior to the
scheduled surgical procedure.

Type of Autologous Donations/Transfusions:

1. Predeposit donation – refers to the blood that is

drawn some time before the anticipated transfusion Red: primary bag
and stored, usually liquid but occasionally frozen (will
2 colorless: satellite bag
expire for 10 years- pinaka matagal)
2. Intraoperative autologous transfusion – occurs Label: fresh frozen plasma (FFP) then Platelet
when blood is collected during the surgical procedure concentrate (PC). May serial number and dates (DOE:
and usually re-infused immediately. date of extraction) (DOExp: Date of expiration
3. Immediate preoperative hemodilution – takes depending on the anticoagulant)
place in the operating room when 1-3 units
of WB are collected and the patient’s volume is
replaced with colloid or crystalloid. The blood is re- Blood Component
infused during the surgical procedure.
4. Post-operative salvage – an autologous donation
in which a drainage tube is placed in the surgical site
Schematic diagram of blood component
and postoperative bleeding is salvaged, cleaned and
re-infused.

- Donor blood testing: need ng 5mL (red (IS): rapid

plasma regain, HCV, HBsAg, HIV) and 5mL (EDTA
(Hema): malarial smear, hemoglobin, hematocrit)
tube. So need mag extract ng 10cc

- How long ang blood collection? range of 5-10

minutes with an average of 7-8 minutes maximum - ang storage ng whole blood sa ref is 1-6’ centigrade
time of allogenic blood collection is 12 minutes (check
- Color coded ang sticker sa blood bag. Kapag A: blue
for clot na)
, B: Yellow , AB: Pink, O- white
- Component preparation: We need to separate the
- Blood bag 150 mL with 63mL anticoagulant
whole blood to plasma and platelet component (18 is
much preferred) - Close system: If the needle is cut before the sealed
area
COMPONENT PREPARATION & INDICATIONS
- Open system: if the cut is after the sealed. It is
PRIOR TO BLOOD COLLECTION, the intended
needed to be check to look for lysis (hemolysis). If
venipuncture
plasma is cut open, yellow green color plasma will be
site must be cleaned with a scrub solution containing:
observed meaning it will be considered as bacterial
a. hypochlorite
infection (Pseudomonas aeruginosa
b. isopropyl alcohol (/) 70%
c. 10% acetone Screening Test
d. PVP iodine complex (/)
18-17/19 gauge (blood bag) - Kusang mamatay sa refrigerated temp si Malaria and
- naka bevel down ang needle (para maiwasaan ang RPR
pagsirit ng dugo) - HBV< HIV< HCV
- tourniquet should be 3 inches away from
venipuncture size

JD 💉
BLOOD COMPONENT PREPARATION - Request: PRBC

- Bakit PRBC lang ang binibigay sa patient na may

anemia? Kasi RBC lang naman ang needed sa anemia,
ang whole blood kasi ay may lamang plasma and baka
magresult pa sa Circulatory Overload si patient since
hindi naman niya yon needed

LEUKOPOOR RED BLOOD CELLS

Shelflife: closed system – same with Packed RBC

Open System – 24 hours

Blood Components Storage Temp: 1-6ºC

Oxygen Carrying Components/Products Contents: 5 x 106 residual WBC

• Red cell concentrates
• Leukocyte-poor red blood cells
• Frozen-thawed red cells
Platelet Products
• Platelet rich plasma (PRP)
• Platelet concentrates (PC)
Plasma Products
• Fresh frozen plasma (FFP)
• Frozen plasma (FP)
• Cryoprecipitate
• Stored plasma
PLASMA DERIVATIVES
Ex. NSA ISG
CONC. PPF Rhogam FACTOR IX CONC.
Indications: Anemia with history of febrile reactions
(kapag nilalagnat during transfusion); to decrease
alloimmunization (alloantibodies has been formed) to
WBC or HLA antigens or CMV transmission
- mababa ang leukocytes
REJUVENATED RED BLOOD CELLS
Addition of Rejuvenation soln ( PIGPA-Phosphate,
Inosine, Glucose, Pyruvate, Adenosine) to regenerate
ATP and 2,3-DPG.
FACTOR VIII Shelflife: can be prepared 3 days after expiration date
Storage Temp: 1-6ºC

Component Transfusion Therapy *For transfusion, wash properly (using

LISS or NSS para maalis yung mga chemicals na hindi
- one unit may be used for multiple transfusion needed) and transfuse within 24 hours.

- it is the effective utilization of a limited natural WASHED RED BLOOD CELLS

resource by providing therapeutic material to several
patients from a single donation Shelflife: Open System: 24 hours

WHOLE BLOOD Storage Temp: 1-6ºC

QC Requirement: Plasma removal

Indications: anemia with history of febrile reactions;

PNH (Paroxysmal nocturnal hemoglobinuria: patient
na lumalabas ang hemoglobin sa urine tuwing gabi);
for pxs with plasma proteins antibodies to reduced
allergic reactions (for IgA-deficient pxs)

- After preparing packed rbc, pwede na itong isunod

- Pinaka common anticoagulant sa bloodbank is CPD
and ACD. Hindi masyadong nagagamit ang heparin
pero nagagamit siya sa emergency transfusion.
- Exchange transfusion ay nangyayari dun sa mga may
Rh incompatibility na baby.
- Kapag mag rrequest sa blood bank, ang nakalagay is
1 “u” FWB (Fresh whole blood)
PACKED RED BLOOD CELLS
Shelflife: same with WB
Storage Temp: 1-6ºC
Contents: Hematocrit should be 80 % or less
FROZEN< THAWED< DEGLYCEROLIZED RBC
Shelflife: Frozen – 10 years (pinaka matagal)
Deglycerolized – 24 hours
Storage Temp: freezing - - 65ºC (High Glycerol-
40%),- 120ºC (Low Glycerol-20%),
- 65ºC (using
79%glycerol with dextrose, fructose and EDTA)
Deglycerolizing Process - 1-6ºC
Indications: anemia, long term storage of “rare”
units and/or autologous units

Indication: Restore oxygen carrying capacity (anemia)

Immediate effect of one unit: increase Hematocrit by

3% and increase hemoglobin by 1g

JD 💉
PLATELETS (RANDOM DONOR, prepared from whole
blood)
Shelflife: 3-5 days (5 days with continuous agitation)
Storage Temp: 20-24ºC with constant agitation
Contents: 5.5 X 1010 platelets in 50-65 mL of plasma
Indications: Thrombocytopenia, Dengue
hemorrhagic fever (kapag random ang blood, need
iprocess immediately), DIC, platelet disorders,
bleeding
Immediate effect: increase platelet count by 5,000-
10,000 per unit
- Agitation machine is used to prevent clumping of
platelet
PLATELETS (SINGLE DONOR, prepared by pheresis)
Shelflife: Closed system – 5 d
Open system - 24 hours
Storage Temp: 20-24ºC with constant agitation
Contents: 3.0 X 1011 platelets in approx. 300 mL of
plasma
Indications: Thrombocytopenia; for pxs refractory to
randomplts. due to platelet antibodies
Immediate effect: increase platelet count by 30,000-
60,000/unit
FRESH FROZEN PLASMA (SINGLE DONOR, prepared
from whole blood)
Shelflife: frozen= 1 year thawed= 24 hours
Storage Temp:frozen= -18ºC thawed= 1-6ºC
Contents: All coagulation factors; 400mg Fibrinogen
Indication: Treatment of multiple coagulation factor
deficiencies (caused by massive transfusion, trauma,
liver dse, DIC)
Also for treatment of AntiThrombin III deficiency, TTP,
HUS
- Bakit plasma eh platelets ang needed nila? Ang
dengue patients, they are hemo concentrated –
mataas ang hematocrit at kapag mataas ang
hematocrit, usually mababa ang platelets
- This is the blood component that doesn’t need
crossmatching. Ang binabasehan is blood type using
reverse typing.
Question:
a. How many units of FFP must be transfused to
increase the fibrinogen level from 200 mg to 1000
mg?
Answer: needed ng 2 units para may 800mg
b. FFP: Date of Collection= Jan. 15, 2008
Thawed @ 10:00am, Feb. 01, 2008
When will be the expiration date?
Answer: February 2, 2008, 10:00am
SINGLE DONOR PLASMA (SDP) LIQUID/FROZEN
Shelflife: Liquid – 5 days beyond whole blood
expiration
Frozen – 5 years
Storage Temp: Liquid – 1-6ºC
Frozen= -18ºC or colder
Indication: Treatment of stable clotting factor
deficiencies
CRYOPRECIPITATED ANTIHEMOPHILIC FACTOR
(CRYOPRECIPITATE)
Shelflife: Frozen – 1 year
Thawed – 6 hours
Pooled – 4 hours (yung 4 unit will be
transferred into 1 unit of blood bag- combination of
more than 1 unit of blood bag)
Storage Temp:Frozen= -18ºC or colder
Thawed – 20-24ºC
Thawed process: pwedeng ilagay sa running water,
pwede sa waterbath at 37’C, or iiwanan lang sa RT
Contents: Factor VIII:C - 80-150 IU
Factor VIII:vWF
Fibrinogen – 150-250mg
Factor XIII
Indications: Hemophilia A, von Willebrand’s dse,
Fibrinogen deficiency, Factor XIII deficiency – all ay
coagulation problems
***FIBRIN GLUE –
GRANULOCYTE CONCENTRATE
Shelflife: 24 hours
Storage Temp: 20-24ºC without agitation
Contents: 1 x 1010 wbc
Indications: To correct severe neutropenia, fever
unresponsive to antibiotic therapy and myeloid
hypoplasia of the bone marrow
PLASMA DERIVATIVES
are concentrates of plasma proteins that are
prepared from pools (many units) of plasma.
JD 💉
FACTOR VIII CONCENTRATE (irradiation uses Cesium-137 or Cobalt-60)

Shelflife: varies on expiration date on vial Indications: GVH reactions, BM trnsplant, direct
donation from a blood relative, exchange transfusion,
Storage Temp: 1-6ºC (lyophilized- it is innate powder
IUT, transfusion for immunocompromised patients
form and nasa refrigerator)

Indication: Hemophilia A

FACTOR IX CONCENTRATE (PROTHROMBIN

COMPLEX)

Shelflife: varies on expiration date on vial

Storage Temp: 1-6ºC (lyophilized)

Indication: Hemophilia B

IMMUNE SERUM GLOBULIN (ISG)

Shelflife: Intramuscular: 3 years

Intravenous: 1 year

Indications: prophylactic treatment to pxs exposed to

hepatitis, measles or chickenpox; treatment of
congenital hypogammaglobulinemia

NORMAL SERUM ALBUMIN (NSA)

Shelflife & Storage Temp: 3 years at 20-24ºC

5 years at 1-6ºC

Contents: 96% Albumin and 4% Globulin

Indications: Plasma volume expansion: surgery,

trauma, burns

- nirready sa emergency cases

PLASMA PROTEIN FRACTION

Shelflife & Storage Temp:3 years at 20-24ºC

5 years at 1-6ºC

Contents: 80-85% Albumin and 15-20% Globulin

Indication: Plasma volume expansion

SYNTHETIC VOLUME EXPANDERS

NSS

Ringer’s lactate

Electrolyte Solution

Dextran

Hydroxyethyl starch (HES)

Rho (D) Ig (Rhogam)

Shelflife and Storage Temp: 3 years at 1-6ºC

Contents: Full dose- 300ug Anti-D

Mini Dose- 50ug Anti-D

Indication: Prevention of Rho (D) immunization

IRRADIATED BLOOD

Shelflife: 28 days or the normal dating period of the

blood, which ever comes first

JD 💉