Beruflich Dokumente
Kultur Dokumente
1667
▫ Jean Baptiste Denis:
▫ First recorded animal-to-human blood transfusion (calf blood)
▫ Richard Lower: Sheep’s blood
1795
▫ Philip Syng Physick:
▫ Unconfirmed first human-to-human transfusion
Historical Overview
1818
▫ James Blundel of England
▫ Successful transfusion to a woman suffering form
postpartum haemorrhage.
1869
▫ Braxton Hicks
▫ First non-toxic anticoagulant: sodium phosphate
Historical Overview
1901
▫ Karl Landsteiner, Discovery of ABO blood groups,
"Specificity of Serological Reactions”
1902
▫ Anthony Decastello and Adriano Sturli
▫ AB blood group
Historical Overview
Edward E. Lindemann
▫ Was first to successfully carry out vein to vein
transfusion of blood by using multiple syringes and a
special cannula for puncturing the vein through the
skin.
▫ Unger designed his syringe-valve apparatus.
Historical Overview
1907
▫ Richard Weil
▫ 1st to perform ABO typing and began compatibility
testing
▫ 1st to suggest ABO inheritance
1913
▫ Reuben Ottenberg stressed importance of
compatibility testing.
Historical Overview
1914
▫ Albert Hustin, sodium citrate as an anticoagulant
solution.
1915
▫ Richard Lewisohn, minimum amount of citrate
needed for anticoagulation.
Historical Overview
1916
▫ Rous and Turner, introduction of citrate dextrose
solution for RBC preservation.
1930’s
▫ Function of glucose in RBC metabolism was known.
1932
▫ First blood bank in Leningrad, Russia.
Historical Overview
1939-1940
▫ Philip Levine (together with Stetson, Landsteiner
and Alex Wiener), 1st discovery of Rh blood groups
1941
▫ Charles Drew developed techniques in blood
transfusion and blood preservation during WWII.
1943
▫ Loutit and Mollison introduced the used of ACD
Historical Overview
1945
▫ Coomb, Mourant, Race, Antihuman globulin reagent (was
first described by Carlo Moreschi in 1908)
1947
▫ Rh immune Globulin for prevention of Hemolytic Disease of the
Fetus and Newborn
1951
▫ Edwin Cohn ,development of cell separator, paved the way for
component therapy.
▫ Carl Walter, blood collection using a collapsible bag of polyvinyl
resin.
Historical Overview
1957
▫ Gibson
▫ Introduction of citrate-phosphate-dextrose (CPD)
1960
▫ Plasmapheresis (therapeutic)
1985
▫ DR. Yves Lapierre developed Gel test in Lyon,
France
Basic Genetics
Genetics
• The study of inheritance or the transmission of
characteristics from parents to offspring.
Levels of Genetics:
• Population, concerning genetic traits in large numbers
of individuals.
• Cellular, which pertains to the cellular organization of
genetic material.
• Molecular, based on the biochemistry of genes and the
structures that support them.
Deoxyribonucleic Acid
Phenotype:
• Anything that is produced by the genotype, including an
enzyme to control a blood group antigen; the length of
long bones of the skeleton etc.
Terminologies
Amorphic gene:
• A gene with no observable effect, manifestation or
product.
Hemizygous:
• Refers to the condition when one chromosome has a
copy of the gene and the other chromosome has that
gene deleted or absent.
Terminologies
Codominant:
• Equal expression of both alleles in phenotype
• Most of the antigens in the various blood group systems
(i.e., ABO,Rh, Kell, Kidd, etc.) generally follow
straightforward inheritance patterns, usually of a
codominant nature.
Antithetical:
• Opposite form of a gene, different allele.
Terminologies
• Cis: Genes are inherited on the same chromosome.
C in cis position to D:
DCe/dce
• Trans: Genes are inherited on separate chromosomes.
Genes inherited in transposition can weaken the trait's
expression.
C in trans position to D:
Dce/dCe
Terminologies
Linked genes
• Genes that are close together on a chromosome and
inherited as one unit.
Haplotype
• Set of genes inherited via one of the two parental
gametes.
Terminologies
Dosage:
If antibody gives a stronger reaction with RBCs double-
dosed for the target antigen, it shows the dosage effect.
Patterns of Inheritance
Autosomal dominant:
▫ Genes expressed with equal frequency in males and females, on
non-sex chromosome.
Sex-linked dominant:
▫ Carried on the X chromosome; no father-to-son transmission;
will be expressed if passed from father to daughter or from
mother to son.
Sex-linked recessive:
▫ Carried on the X chromosome.
▫ Males inherit it from carrier mothers; traits are exhibited most
commonly in males. (e.g., hemophilia A).
▫ Females can exhibit the trait but must inherit it from both carrier
mother and affected father.
Mendelian Inheritance Principles
Law of Independent Segregation
▫ Two members of a single gene pair passed from one
generation to the next in separate gametes.
Law of Dominance
▫ Recessive alleles will always be masked by dominant
alleles.
Law of Independent Segregation
Law of Independent Assortment
Hardy-Weinberg Principle
Mathematical formula that allowed the study of Mendelian
inheritance in great detail.
Hardy-Weinberg formula:
p+q=1
• Acquired or adaptive
▫ The specific, evolved IS
Humoral, mediated by B cells and antibody
production
Cellular, mediated by T cells and lymphokines
Characteristics of Antigens
• Antigens are substances that combine with an
antibody.
• An antigen that causes a specific immune response is an
immunogen.
• 23 RBC antigen systems containing over 200 RBC
antigens.
Proteins -Rh, M, and N
Glycolipids - ABH, Lewis, Ii, and P
Glycoproteins - HLA
Antibody Characteristics
Characteristics of Blood Group Antibodies
Naturally Occurring Immune Antibodies
• Found in the serum of • Found in the serum of
individuals who have never individuals who have been
been previously exposed to RBC transfused or who are pregnant.
antigens by transfusion,
injection, or pregnancy.
Public Antigen
▫ Antigen found commonly among individuals
▫ More than 98% of the population
General characteristics
Low-Frequency antigen
▫ “Low-incidence”
▫ Prevalence of less than 1% of most random
populations
High-Frequency antigen
▫ “High-incidence”
▫ Prevalence of more than 90% of most random
populations
Summary of Blood Group Systems
ISBT NUMBER BLOOD GROUP CHROMOSOME NO.
001 ABO 9q
002 MNS 4q
003 P 22q
004 Rh 1p
005 Lutheran 19q
006 Kell 7q
007 Lewis 19p
008 Duffy 1q
009 Kidd 18q
010 Diego 17q
Summary of Blood Group Systems
ISBT NUMBER BLOOD GROUP CHROMOSOME NO.
011 Cartwright 71
012 Xga Xp
013 Scianna 1p
014 Dombrock 12p
015 Colton 7p
016 Landsteiner-Wiener 19p
017 Chido/Rodgers 6p
018 H 19q
019 Kx Xp
020 Gerbich 2q
Summary of Blood Group Systems
ISBT NUMBER BLOOD GROUP CHROMOSOME NO.
021 Cromer 1q
022 Knops 1q
023 Indian 11p
024 Ok 19p
025 Raph 11p
026 John-Milton Hagen 15q
027 I 6p
028 Globoside 3q
029 GIL 9p
030 RHAG 6p
ABO blood group system
ABO blood group system
• The ABO system is the most important of all blood
groups in transfusion practice.
• Saliva
• Tears
• Urine
• Digestive juices
• Bile
• Milk
• Amniotic fluid
• Pathological fluids:
▫ Pleural
▫ Peritoneal
▫ Pericardial
▫ Ovarian cyst
Secretor Status
Procedure:
1. Collect 2 to 3 ml saliva in a clean 16 x 100 mm tube.
Use paraffin wax or clean rubber bands to stimulate
secretions.
2. Centrifuge at 1000xg for 8 to 10 minutes
3. Transfer supernatant in a stoppered tube.
4. Place in a boiling waterbath for 10 minutes. This
inactivates enzymes that might otherwise destroy blood
group substances.
5. Recentrifuge and collect clear supernatant.
6. Dilute saliva with NSS.
Secretor Status
8. Add one drop of the appropriate diluted antiserum to
each tube.
9. Add one drop of supernatant saliva.
10. Mix and incubate for 8 to 10 minutes.
11. Add one drop of appropriate indicator cells.
12. Mix and incubate at RT for 30-60 mins
13. Centrifuge
14. Observe for macroscopic agglutination
Secretor Status
SAMPLE: Saliva
PRINCIPLE: Agglutination Inhibition
+ Agglutination: Negative
- Agglutination: Positive
A
A
A A A
A Y Y
A
A
A A A
Y Y
A
A SeSe Y Indicator Cells
Anti-A
Secretor Status
SAMPLE: Saliva
PRINCIPLE: Agglutination Inhibition
+ Agglutination: Negative
- Agglutination: Positive
No agglutination:
A A Secretor
Y A Y
A A
A SeSe Y A
Anti-A
ABO Subgroups
A Subgroups
A1 Versus A2 Phenotypes
Blood group Anti-A Anti-A1 Lectin
(anti-A plus anti-A1)
A1 + +
A2 + 0
O O O + + +
Oh
Group O vs Oh
Group Anti A Anti B Anti H A cells B cells
O O O + + +
Oh O O O + +
Para-Bombay phenotype
• RBCs are completely devoid of H antigens or have small
amounts of H antigen present.
Universal Recipient:
Universal Recipient: AB
0 0 + + 0
+ 0 0 + A
0 + + 0 B
+ + 0 0 AB
ABO Discrepancies
• ABO discrepancies occur when the forward and reverse
groupings do not agree.
Rouleaux
Group III Discrepancies
• Elevated levels of globulin from certain diseases:
▫ Multiple myeloma
▫ Waldenström’s macroglobulinemia
• Hodgkin’s lymphomas
• Elevated levels of fibrinogen
• Plasma expanders (dextran & polyvinylpyrrolidone)
• Wharton’s jelly in cord blood samples
Group III Discrepancies
Example:
Forward Typing Reverse Typing
Anti-A Anti-B A1 cells B cells
4+ 2+ 4+ 2+
Group III Discrepancies
Resolution:
• Washing the patient’s red cells with saline or adding a
drop or two of saline to the tube in case of rouleaux
formation.
1 2
D C E
Wiener Fisher-Race Wiener Fisher-Race
R1 DCe r’ dCe
R2 DcE r’’ dcE
Ro Dce r dce
Rz DCE ry dCE
Convert:
R1r” to Fisher-Race:
R2Rz to Fisher-Race:
Convert:
R1r” to Fisher-Race:
DCe/dCe
R2Rz to Fisher-Race:
DcE/DCE
Rosenfield and Coworkers:
Alphanumeric Terminology
D + C + E + c negative, e negative
Rosenfield
• D= Rh1
• C= Rh2
• E= Rh3
• c= Rh4
• e= Rh5
D + C + E + c negative, e negative
Rh: 1, 2, 3, –4, –5
International Society of Blood Transfusion
Committee
• Six-digit number for each authenticated antigen
belonging to a blood group system
Example:
Dce/dCe
Weak D
• Results from inheritance of RHD genes that code for a
weakened expression of the D antigen
• D antigens are complete but fewer in number
Amorphic
▫ RHD gene is absent, no expression of RHCE gene
Regulator
▫ Gene inherited, but not expressed
Rh Deficiency Syndrome
Rh mod phenotype
• Partial suppression of RH gene expression caused by
mutations in the RHAG gene.
• RhAG protein is altered, normal Rh antigens are also
altered, often causing weakened expression of the
normal Rh and LW antigens.
• Exhibit features similar to those with the Rh null
syndrome.
• Clinical symptoms are usually less severe.
Unusual phenotypes
Cw
▫ Results in a single amino acid change most often found
on the RhCe protein.
▫ Found in about 2% of whites and is very rare in blacks.
▫ Anti-Cw has been identified in individuals without
known exposure to foreign RBCs and after transfusion
or pregnancy.
▫ Anti-Cw may show dosage
Unusual phenotypes
f (ce)
▫ Expressed on the RBC when both c and e are present
on the same haplotype.
▫ Compound antigen
▫ Anti-f only shows postive reactivity with DCE/dce.
▫ Anti-f is generally a weakly reactive antibody often
found with other antibodies.
Unusual phenotypes
rhi (Ce)
▫ Compound antigen present when C and e are on
the RhCe protein.
• DAT +
• Antibody screen (+) (-)
• Antibody elution
Rh HDFN
• Severe
• Rh antibodies are primarily IgG, which readily cross the
placenta
• Mother:
• Father:
• Baby:
Rh HDFN
• Severe
• Rh antibodies are primarily IgG, which readily cross the
placenta
• Mother: Rh Negative
• Father: Rh Positive
• Baby: Rh Positve
Other Blood group Systems
The Lewis (007) System
• Lewis antigens are not intrinsic to RBCs but are on type 1
glycosphingolipids that are passively adsorbed onto the
RBC membrane from the plasma.
GlcNAc R
GAL • Lipid
• Protein
• Secretions - glycoproteins.
• Plasma - glycolipids.
Se allele (FUT2) Secretor Enzyme
SeSe
Sese
GlcNAc R
GAL
FUC Type 1 H
Le allele (FUT3) Lewis Enzyme
LeLe
Lele FUC
GlcNAc R
GAL
Le a
Interaction between Le and Se enzymes
FUC
GlcNAc R
GAL
FUC
Le b
Lewis enzyme substrates
Substrate Antigen
Type 1 Lea
Type 1 H Leb
Type 1 A ALeb
Type 1 B ALeb
Phenotypes of the Lewis System
Le and Se genes inherited
Le(a-b-) Le(a+b-)
• Types:
▫ Anti-Lea
▫ Anti-Leb
Lewis antibodies
Anti-Lea
• Most commonly encountered of the Lewis antibodies
• Destroyed by enzymes
▫ Methionine defines S
▫ Threonine defines s.
U-phenotype
• U for UNIVERSAL
• Demonstrate Dosage
• Destroyed by enzymes
• Demonstrate dosage
• Anti-Nf
• Found in renal patients who are dialyzed on equipment
sterilized with formaldehyde.
MNSs Antibodies
Anti-Ss
• IgG, reactive at 37°C and the antiglobulin test
• Destroyed by enzymes
• Auto-anti-i
• Associated with infectious mononucleosis
• Less often a problem than auto-anti-I
• i adult phenotype
• Cataracts
• HEMPAS
The P Blood Group:
P1PK (003), Globoside (028), Related (209) Antigens
• Show dosage
• Antiglobulin reactive.
Summary
Antibody Reactivity Enzyme Treatment
MN RT Destroyed
Ss AHG Variable effect
U AHG No effect
Anti-P1 RT Enhanced
Ii RT Enhanced
Kell AHG No effect
Duffy AHG Destroyed
Lewis Most RT, some Enhanced
37˚C,AHG
LuaIgM RT Variable effect
LubIgG AHG
Kidd AHG Enhanced
MINOR BLOOD GROUP SYSTEMS
The Diego (010) System
• Used as a tool in anthropologic studies of Mongolian
ancestry.
• Composed of 22 antigens.
• Three sets of independent pairs of antithetical antigens:
Dia/Dib Wra/Wrb Wu/DISK
• Anti-Wra
• Some are directly agglutinating, but most require IAT.
• Has caused severe HTRs
• Autoanti-Wrb
• Common in the serum of patients with WAIHA.
The Yt (011) System
• Cartwright, “why T” became “Yt”
• Located on CD44
• Apheresis
– process wherein blood is withdrawn from the donor and
separated into its components
– one or more components is retained and the remaining
constituents are returned
General Requirements for Donation
• General appearance:
▫ Observe the prospective donor for presence of
excessive anxiety, drug or alcohol influence, or
nervousness.
• Age:
▫ at least 17 years old
General Requirements for Donation
• Weight:
▫ At least 110 lbs/50 kg,
▫ Maximum volume of 525 mL can be collected
▫ Maximum of 10.5 mL of blood/kg of donor weight for
WB donation
• Pulse:
▫ Between 50 and 100 bpm
▫ Athletes will have a pulse less than 50 bpm
• Blood pressure:
▫ Less than or equal 180/100 mm Hg
General Requirements for Donation
Hemoglobin:
A. Allogeneic Donation
greater than or equal to 12.5 g/dL
B. Autologous Donation
greater than or equal to 11 g/dL
General Requirements for Donation
Hematocrit:
A. Allogeneic Donation
greater than or equal to 38%
B. Autologous Donation
greater than or equal to 33%
Requirements for allogeneic donation
(Philippine standards)
• Age:
• 16 – 65 years old
▫ (16-17 yrs old requires parent’s consent)
• Temperature:
▫ ≤ 37.5˚c or ≤ 99.5 ˚F
• Pulse rate:
▫ 60-100 bpm, <50 if athlete
• BP:
▫ Systolic 90 to 160 mm Hg
▫ Diastolic 60 to 100 mm Hg
BLOOD COLLECTION PROCESS
1. Donor Registration.
A YELLOW A BLUE
B PINK B YELLOW
AB WHITE AB PINK
O BLUE O WHITE
Donor deferral
Types of deferral
1. Temporary Deferral:
▫ Prospective donor is unable to donate blood for a
limited period of time.
▫ EXAMPLE:
Donor has received a blood transfusion; defer for 12
months from date of transfusion.
Donor deferral
2. Indefinite Deferral:
▫ Prospective donor is unable to donate blood for
someone else for an unspecified period of time due to
current regulatory requirements. These donors may be
eligible to donate autologous blood.
▫ EXAMPLE:
Donor states they have lived in England for 1 year in
1989; defer indefinitely.
Donor deferral
3. Permanent Deferral:
▫ Prospective donor will never be eligible to donate
blood for someone else. These donors may be eligible
to donate autologous blood.
▫ EXAMPLE:
Donor states that he or she has hepatitis C; defer
permanently.
Donor deferral
• PERMANENT DEFERRAL:
▫ Confirmed positive test for HBsAg
▫ Viral hepatitis after 11th birthday
▫ Repeat reactive test for anti-HBc on more than one
occasion
▫ Previous donation associated with hepatitis, HIV or
HTLV transmission
▫ Confirmed positive for HCV, HIV or HTLV
Donor deferral
• PERMANENT DEFERRAL:
▫ Behavioral risk factors for HIV infection
▫ Male-to-male sexual contact even once, since 1977
▫ Men or women who engage in sex for money or drugs
since 1977 are permanently deferred
▫ IV drug users/ Parenteral drug use
▫ Growth Hormone from Human Pituitary Glands
▫ History of Babesiosis or Chaga’s disease
▫ Intake of drug Tegison or etretinate
▫ Hematologic malignancies
Donor deferral
• 1 year deferral:
▫ Blood transfusion
▫ Transplant such as organ, tissue, or bone marrow; or a
graft such as bone or skin
▫ Mucous membrane exposure to blood, non sterile skin
or needle penetration
▫ Sexual contact with anyone who has HIV/AIDS or has
had a positive test for HIV/AIDS
▫ Sexual contact with a prostitute or anyone else who
takes money or drugs or other payment for sex
Donor deferral
• 1 year deferral:
▫ Rape victim
▫ Incarceration in a correctional institution for longer
than 72 hours
▫ History of syphilis or malaria
▫ Sexual contact or living with a person (“close contact”)
who has acute or chronic hepatitis B
▫ History of syphilis or gonorrhea
▫ Travelers to areas the CDC considers to be endemic for
malaria
▫ Hepatitis B Immune Globulin (HBIG)
Donor deferral
• 2-week deferral:
▫ Measles (rubeola)
▫ Mumps
▫ Oral polio
▫ Typhoid
▫ Yellow fever
• 4-week deferral:
▫ German measles (rubella) or chickenpox
▫ Proscar, Propecia and Accutane
Donor deferral
• 12-24 hours:
▫ After recent alcohol intake
• 3 days:
▫ Piroxicam, aspirin (platelet pheresis)
• 6 weeks:
▫ Pregnancy
• 6 months:
▫ Avodart
• 3 years:
▫ Malaria confirmed diagnosis
Autologous donation
“The safest blood that you can receive is your own”
General requirements:
1. Age: no age requirement
2. Hgb :11g/dl, Hct :33%
3. General condition:
Patient should have no condition predisposing to
bacteremia or any form of severe
cardiovascular/pulmonary condition.
4. Single unit is removed at a time, with at least 3 day
intervals, and the final phlebotomy should be at least
72 hours before the surgery.
Types of Autologous Donation
1. Preoperative Collection (Predeposit)
▫ Occurs during the 5 to 6 weeks immediately preceding a
scheduled, elective surgical procedure.
Types of Autologous Donation
2. Acute Normovolemic Hemodilution
▫ Collection of whole blood with the concurrent infusion of
crystalloid or colloid solutions.
▫ 3:1 for crystalloids and 1:1 for colloids
Types of Autologous Donation
3. Intraoperative collection
▫ Involves collecting shed blood from the surgical site, a
device that utilizers vacuum is used to collect shed blood.
▫ Blood is the washed with saline and then concentrated to
reach hematocrit of 50 – 60%, then reinfusing those cells
immediately.
Types of Autologous Donation
4. Postoperative Collection
▫ Collected from a drainage tube placed at the surgical site.
▫ It is reinfused, with or without processing, via a
microaggregate filter to screen out any debris.
Directed Donation
A directed donation is a unit collected under the
same requirements as those for allogeneic donors,
except that the unit collected is directed toward a
specific patient.
Apheresis Donation
• From the ancient Greek aphairesis, “a taking
away”)
• An effective mechanism for collecting a specific
blood component while returning the remaining
whole blood components back to the patient.
Apheresis Donation
Methods of Centrifugation:
1. Intermittent Flow Centrifugation
▫ Blood is processed in batches or cycles
▫ Requires only one venipuncture
▫ Blood is drawn and reinfused through the same needle
▫ Once the desired component is separated, the
remaining components are reinfused to the donor.
Apheresis Donation
2. Continuous flow centrifugation (CFC)
▫ Two venipuncture sites are necessary
▫ Involves withdrawal and processing and reinfusing of
blood to the individual simultaneously
Apheresis Donation
• Amount of time for a particular procedure can range
from 45 to 120 minutes.
Management:
1. Remove the tourniquet and withdraw needle
2. Place cold compresses on the donor’s forehead
3. Raise the donor’s legs above the level of the head
4. Loosen tight clothing and secure airway
5. Monitor vital signs
Donor Reactions
Mild Reactions:
Twitching or muscle spasms
Management:
Disengage the hyperventilation sequence by conversing
with the donor and having the donor breathe into a paper
bag.
Donor Reactions
Mild Reactions:
Nausea or vomiting
Management:
1. Instruct the donor to breathe slowly
2. Apply cold compresses to the forehead
3. Turn the donor’s head to one side and provide an
appropriate receptacle
4. The donor may be given water after vomiting has ceased
Donor Reactions
Moderate reactions:
Loss of consciousness. The donor may have a decreased
pulse rate, may hyperventilate, and may exhibit a fall in
systolic pressure to 60 mm Hg.
Management:
1. Check vital signs frequently
2. Administer 95% oxygen and 5% carbon dioxide
Donor Reactions
Severe reactions:
Convulsions can be caused by cerebral ischemia, marked
hyperventilation, or epilepsy.
Management:
1. Call for help immediately; notify blood bank physician
2. Try and restrain the donor to prevent injury to self or
others
3. Ensure an adequate airway
Donor Reactions
Severe reactions:
Cardiac or respiratory difficulties
Management:
Perform CPR until medical help arrives
Donor Reactions
Hematomas caused by the needle going through the
vein, with subsequent leakage of blood into the tissue.
Management:
1. Remove the tourniquet and needle from donor’s arm
2. Apply pressure with sterile gauze pads for 7 to 10
minutes, with the donor raising his or her arm above the
heart
3. Apply ice to the area for 5 minutes
Blood preservation and Banking
Blood preservation and Banking
RBC Biology and Preservation:
Three areas of RBC biology are crucial for normal
erythrocyte survival and function:
1. Normal chemical composition and structure of the
RBC membrane
2. Haemoglobin structure and function
3. RBC metabolism
The RBC’s metabolic pathways that produce ATP are
mainly anaerobic.
Citrate-Phosphate-Dextrose CPD 21
Citrate-Phosphate-Double-Dextrose CD2D 21
• Additives contain:
▫ Saline
▫ Adenine
▫ Glucose
▫ Mannitol (protects against storage-related hemolysis).
Additive Solutions
Name Abbreviation Storage Time (days)
Adsol AS-1 42
Nutricel AS-3 42
Optisol AS-5 42
Freezing and Rejuvenation
• RBC Freezing is primarily used for autologous units
and the storage of rare blood types.
• Rejuvesol contains:
Phosphate
Inosine
Pyruvate
Adenine
Blood components
Terms to know:
• Whole blood:
Blood collected before separation into components
• Components:
Parts of whole blood that are separated
• Closed system:
A sterile system of blood collection
• Open system:
When the collection is exposed to air, decreasing
expiration date
WHOLE
BLOOD
Heavy spin Light spin
5000 x g for 5 3200 x g for 2-3
mins. mins.
Plasma
Cryoprecipitate
Cryoprecipitate
Whole Blood
Indications Volume expansion, ↑ O2
for use
Storage 1- 6˚c
Temperature
Shelf life • ACD, CPD or CP2D = 21 days
• CPDA1= 35 days
Dosage ↑ hgb 1g/dL
↑ hct 3%
Volume 450–500 mL
Quality Hct approx. 40%
control
Packed RBCs
Indications for Symptomatic anemia with normal blood
use volume, ↑ O2
Storage 1- 6˚c
Temperature
Shelf life • Open system= 24 hours
• ACD, CPD or CP2D = 21 days
• CPDA1= 35 days
• AS=42 days
Dosage ↑ hgb 1g/dL
↑ hct 3%
Volume 250-300 ml
Quality control Hct. ≤80%
Washed RBCs
Indications Symptomatic anemia on patients with
for use severe allergic or anaphylactic conditions
(IgA-negative persons)
Storage 1- 6˚c
Temperature
Shelf life 24 hours
Dosage ↑ hgb 1g/dL
↑ hct 3%
Volume 180 mL
Quality Hct 70–80%
control
RBC leukoreduced
Indications Prevents febrile transfusion reaction and
for use TRALI caused by Abs againts WBC,
Prevent CMV
Storage 1- 6˚c
Temperature
Shelf life Closed system: Same
Open system: 24 hours
Dosage ↑ hgb 1g/dL
↑ hct 3%
Volume 250–300 mL
Quality • 5 × 10⁶WBCs
control • ≥ 85% RBC recovery
RBC irradiated
Indications Prevent GVHD, ↑ O2
for use
Storage 1- 6˚c
Temperature
Shelf life Original outdate or 28 days from
irradiation
Dosage ↑ hgb 1g/dL
↑ hct 3%
Volume 250-300ml
Quality 25 Gy to center of canister
control
Irradiated blood components
• Irradiation inhibits the proliferation of T cells and subsequent
transfusion-associated graft-versus-host disease.
Platelet Rich
Heavy spin
Plasma
3600 x g for 5
mins.
Allow the platelets to rest
undisturbed for 1 to 2 hours
at 20°C to24°C
Platelet
Concentrate
• 40 to 70 mL plasma
• Stored at 20°C to 24°C with
continuous agitation
Single Donor Platetet
Indications Platelet Refractoriness, Prevention of
for use HLA alloimmunization
Storage 20–24°C
Temperature
Shelf life 5 days
Dosage ↑30k–60k/μL
Volume 200–400 mL
Quality ≥ 3 × 10¹¹
control pH ≥ 6.2
Refractory Patients
• Massive splenomegaly
• High fever
• Sepsis
• DIC
• Platelet or HLA antibodies
Delayed transfusion
Transfusion Reactions
• Acute/ Immediate transfusion reaction
Transfusion reaction with signs or symptoms presenting
during or within 24 hours of transfusion.
• Citrate toxicity
Transfusion Transmitted Diseases
Disease Transmission Prevention—Required Tests
Diseases REQUIRED TESTS
Hepatitis B HBsAg, anti-HBc
Hepatitis C Anti-HCV, HCV RNA
HIV Anti-HIV-1/2 HIV-1 RNA
HTLV Anti-HTLV-I/II
Syphilis STS
West Nile Virus WNV RNA
Other Viruses
• Cytomegalovirus
Most frequently transmitted virus from mother to
fetus.
ELISA, FIA, indirect hemagglutination.
• Epstein-Barr Virus
• Kissing disease
• Infectious mononucleosis
Other Viruses
• Parvovirus B19
“Fifth disease”
Enters the RBC via P antigen & replicates in the
erythroid progenitor cells
PCR
• HHV-6
• Roseola infantum or sixth disease.
• blood components are not being tested for HHV-6
• HHV-8
• Kaposi’s sarcoma (KS), primary effusion lymphoma
• No evidence to support a TTD association
Transfusion-Associated Parasites
• At least three parasites have been associated
with transfusion associated infections:
▫ Babesia microti
▫ Trypanosoma cruzi
▫ Malaria (Plasmodium species)
Bacterial Contamination
• Incidence of transfusion-associated bacterial sepsis is
low, the morbidity and mortality rates are high.
• Common sources of bacterial contamination include
donor skin and blood.
• Platelets have been the most frequent source of septic
transfusion reactions.
• According to the CDC, Yersinia enterocolitica is the
most common isolate found in RBC units, followed by
the Pseudomonas species.
Bacterial Contamination
• Propionibacterium acnes, a common isolate of human
skin, was the most common bacterial contaminant in
RBCs.
Lattice formation
Basic Red Cell- Antibody Interactions
Hemolysis
• Direct lysis of RBCs due to antibody coating and
complement activation.
• Uncommon, but equal to agglutination.
• Usually due to IgM antibodies.
Factors affecting antigen-antibody reaction
• Temperature
▫ Cold-reactive RT or colder (IgM)
▫ Warm-reactive 37 deg C (IgG)
▫ Must react in appropriate temperature for best
antibody detection.
• pH
▫ Optimal pH 6.5-7.5 or 7
▫ Antibodies enhanced by acidic pH (6.5):
Anti-D, Anti-M, anti-Pr
Factors affecting antigen-antibody reaction
▫ 133:1
▫ (4 drops of serum with 1 drop of a 3% RBC solution)
Factors affecting antigen-antibody reaction
Factors affecting antigen-antibody reaction
• Incubation time
• Incubation times may vary between 30 and 120
minutes.
Anti-IgG
Gel Testing
Anti-IgG
Gel Testing
Serum
Anti-IgG
Gel Testing
Anti-IgG
NEGATIVE
Gel Testing
Gel Testing
Solid Phase Adherence Method
No patient Ab present
Solid Phase Adherence Method
Solid Phase Adherence Method
Antiglobulin Test
• Antihuman globulins (AHGs) obtained from
immunized nonhuman species bind to human globulins
such as IgG or complement, either free in serum or
attached to antigens on red blood cells.
• Monospecific AHG:
▫ Anti-IgG or antibody to anti–C3b-C3d.
Ask the patient to state his or her full name and to spell it
out.
Collection of Patient Samples
• Blood samples should be drawn, carefully using a
technique that avoids hemolyzing the sample.
• Group O cells are used so that anti-A and anti-B will not
interfere in the detection of antibodies to other blood
group systems.
Antigen profile sheet
• Ideally, there will be homozygous expression of many
of the antigens within the screen cell set, allowing for
detection of antibodies that show dosage.
Autocontrol
Patient RBCs
+
Patient serum
• IS
3+ o o
o o o
o o o
3+ o o
o o o
o o o
3+ o o
o o o
3+ o o
o o o
o o o
o o o
Interpreting Antibody Panels
Ruling out:
• Negative reaction (0) indicates that the antibody(ies)
does(do) not react with any antigen on that RBC.
\ 3+ o o
o o o
o o o
3+ o o
o o o
o o o
3+ o o
o o o
3+ o o
o o o
o o o
o o o
Ruling out
3+ o o
o o o
o o o
3+ o o
o o o
o o o
3+ o o
o o o
3+ o o
o o o
o o o
o o o
Ruling out
3+ o o
o o o
o o o
3+ o o
o o o
o o o
3+ o o
o o o
3+ o o
o o o
o o o
o o o
Ruling out
3+ o o
o o o
o o o
3+ o o
o o o
o o o
3+ o o
o o o
3+ o o
o o o
o o o
o o o
Ruling out
3+ o o
o o o
o o o
3+ o o
o o o
o o o
3+ o o
o o o
3+ o o
o o o
o o o
o o o
Ruling out
3+ o o
o o o
o o o
3+ o o
o o o
o o o
3+ o o
o o o
3+ o o
o o o
o o o
o o o
Ruling out
3+ o o
o o o
o o o
3+ o o
o o o
o o o
3+ o o
o o o
3+ o o
o o o
o o o
o o o
Ruling out
3+ o o
o o o
o o o
3+ o o
o o o
o o o
3+ o o
o o o
3+ o o
o o o
o o o
o o o
Ruling out
3+ o o
o o o
o o o
3+ o o
o o o
o o o
3+ o o
o o o
3+ o o
o o o
o o o
o o o
Ruling out
3+ o o
o o o
o o o
3+ o o
o o o
o o o
3+ o o
o o o
3+ o o
o o o
o o o
o o o
Ruling out
3+ o o
o o o
o o o
3+ o o
o o o
o o o
3+ o o
o o o
3+ o o
o o o
o o o
o o o
Ruling out
3+ o o
o o o
o o o
3+ o o
o o o
o o o
3+ o o
o o o
3+ o o
o o o
o o o
o o o
Ruling out
3+ o o
o o o
o o o
3+ o o
o o o
o o o
3+ o o
o o o
3+ o o
o o o
o o o
o o o
Ruling out
3+ o o
o o o
o o o
3+ o o
o o o
o o o
3+ o o
o o o
3+ o o
o o o
o o o
o o o
Ruling out
3+ o o
o o o
o o o
3+ o o
o o o
o o o
3+ o o
o o o
3+ o o
o o o
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o o o
Ruling out
3+ o o
o o o
o o o
3+ o o
o o o
o o o
3+ o o
o o o
3+ o o
o o o
o o o
o o o
Ruling out
3+ o o
o o o
o o o
3+ o o
o o o
o o o
3+ o o
o o o
3+ o o
o o o
o o o
o o o
Ruling out
3+ o o
o o o
o o o
3+ o o
o o o
o o o
3+ o o
o o o
3+ o o
o o o
o o o
o o o
Ruling out
3+ o o
o o o
o o o
3+ o o
o o o
o o o
3+ o o
o o o
3+ o o
o o o
o o o
o o o
Ruling out
3+ o o
o o o
o o o
3+ o o
o o o
o o o
3+ o o
o o o
3+ o o
o o o
o o o
o o o
Ruling out
3+ o o
o o o
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3+ o o
o o o
o o o
3+ o o
o o o
3+ o o
o o o
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Ruling out
3+ o o
o o o
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3+ o o
o o o
o o o
3+ o o
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3+ o o
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o o o
Ruling out
3+ o o
o o o
o o o
3+ o o
o o o
o o o
3+ o o
o o o
3+ o o
o o o
o o o
o o o
Circle antigens not crossed out
3+ o o
o o o
o o o
3+ o o
o o o
o o o
3+ o o
o o o
3+ o o
o o o
o o o
o o o
Look for a matching pattern
3+ o o
o o o
o o o
3+ o o
o o o
o o o
3+ o o
o o o
3+ o o
o o o
o o o
o o o
Look for a matching pattern
3+ o o
o o o
o o o
3+ o o
o o o
o o o
3+ o o
o o o
3+ o o
o o o
o o o
o o o
Anti-Lea
Guidelines:
Autocontrol:
• Negative: Alloantibody
• Positive: Autoantibody or DTR
Phases:
• IS – Cold (IgM)
• 37˚ - Warm reacting (IgM or IgG)
• AHG – Warm (IgG)
Reaction strength:
• Single-strength reactions usually indicate a single antibody.
• Single antibody:
▫ If there is only one antibody, the reactions will match the
antigen pattern on the antigram.
• Multiple antibodies:
▫ If there is more than one antibody, the reactions are
difficult to match with a single antigen pattern on the
antigram.
Rule of three
• The rule of three must be met to confirm the presence
of the antibody
• A p-value ≤ 0.05 must be observed
• This gives a 95% confidence interval
• How is it demonstrated?
Patient serum MUST be:
Positive with 3 cells with the antigen
Negative with 3 cells without the antigen
3+ o o
o o o
o o o
3 Positive 3+ o o
cells o o o
o o o
3+ o o
o o o
3 Negative 3+ o o
cells o o o
o o o
o o o
Additional Techniques for Antibody Identification
• For example:
#5 0 + 0 0 0 3+
#8 0 0 + 0 0 0
Enzymes:
• Ficin (Figs) – most common
• Papain (Papaya)
• Bromelin (Pineapple)
• Trypsin (calf spleen)
Enzymes
M M M
Enzymes
M M M
Enzymes
M M M
Enzymes
Y Y
M M M
Enzymes
Y Y
M M M
Enzymes
Y Y
M M M
Enzymes
Y Y
Enzymes
Enzymes modify the RBC surface by removing sialic acid
residues and by denaturing or removing glycoproteins.
Enhanced Destroyed
Rh Duffy
Kidd MNS
Lewis Xga
P1 Ch/Rg
I
ABO
Enzyme techniques
• One-step
▫ Enzyme is added directly to the serum/cell
mixture
• Two-step
▫ Panel cells are pre-treated with enzyme, incubated
and washed
▫ Patient serum is added to panel cells and tested
Neutralization
Other substances in the body and in nature have antigenic
structures similar to RBC antigens.
Antibody Neutralizing
The patient’s Serum
identification panel substance inhibits
is first incubated
is performed using reactions
with the neutralizing
the between the antibody
substance.
treated serum. and panel RBCs.
Note:
Use of a control (saline and serum) is necessary to prove that the loss
of reactivity is due to neutralization and not to dilution of antibody
strength by the added substance.
Sources of Substances for Neutralization of Certain
Antibodies
ANTIBODY SOURCE OF NEUTRALIZING
SUBSTANCE
Anti-P1 Hydatid cyst fluid, pigeon
droppings, turtledoves’ egg whites
Anti-Lewis Plasma or serum, saliva
Anti-Chido, anti-Rodgers Serum (contains complement)
Anti-Sda Urine
Anti-I Human breast milk
Prewarming
• Performing pretransfusion testing with all reagents and
samples incubated and kept at 37 C.
• Autoadsorption:
▫ Using the patient’s own RBCs to remove autoantibodies.
• Alloadsorption:
▫ Using selected non-self RBCs to remove alloantibodies.
Autoadsorption
Autoantibody
Autoadsorption
Autoantibody
ANTI-K
Autoadsorption
PT PT
K- K-
Autoantibody
ANTI-K
Autoadsorption
Autoantibody
PT
K-
Autoantibody
Autoantibody
PT
ANTI-K K-
Autoadsorption
Autoantibody
PT
K-
Autoantibody
PT
ANTI-K
K-
Alloadsorption
ANTI-K
ANTI-C
ANTI-S
Alloadsorption
ANTI-K K+C+S-
ANTI-C K+C+S-
ANTI-S K+C+S-
Alloadsorption
ANTI-K
K+C+S-
ANTI-C
ANTI-K
K+C+S-
ANTI-C
K+C+S- ANTI-S
Elution
• Removing or “dissociating” an antibody that is attached to the
surface of a red blood cell.
• Methods:
▫ Heat elution
▫ Freeze thaw
▫ Acid (Glycine-HCL-EDTA, digitonin)
▫ Organic solvents (chloroform, Xylene, ether)
Alloadsorption
ANTI-K
K+C+S-
ANTI-C
K+C+S-
Alloadsorption
K+C+S- ANTI-K
TREAT
K+C+S- ANTI-C
Sulfhydryl Reagents
Dithiothreitol (DTT) or 2-mercaptoethanol (2-ME)
• Purpose:
Dissociation of IgG molecules from the surface of
sensitized RBCs and alters the surface antigens of
RBCs.
Chloroquine
• Removes IgG from coated (DAT-positive) RBCs to allow
for accurate phenotyping.
Formula:
___________Number of units needed by the patient______________
Frequency of negativity of antigen #1 x Frequency of negativity of antigen #2
Number of units to crossmatch
Solution:
___2 units____
0.23 x 0.90
= 9.66
• Previous pregnancy
• Transfusion
• During the second and third trimester of pregnancy
Rh HDFN
• Most severe
• Rh antibodies are primarily IgG, which readily cross the
placenta
• Mother:
• Father:
• Baby:
Rh HDFN
• Most severe
• Rh antibodies are primarily IgG, which readily cross the
placenta
• Mother: Rh Negative
• Father: Rh Positive
• Baby: Rh Positve
Hemolysis, Anemia, and Erythropoiesis
Hemolysis
Anemia
erythroblastosis fetalis
Hepatosplenomegaly
hydrops fetalis
• RBC destruction releases hemoglobin, which is
metabolized to bilirubin.
• Phototherapy
▫ Phototherapy at 460 to 490 nm is used to change the
unconjugated bilirubin to isomers, which are less lipophilic
and less toxic to the brain.
Immune hemolytic anemia
Immune hemolytic anemia
Shortened RBC survival mediated through the
immune response, specifically by humoral
antibody.
Three categories:
1. Alloimmune
2. Autoimmune
3. Drug-induced
Immune hemolytic anemia
Alloimmune
Patient produces antibodies to foreign or non-self
RBC antigens introduced into the circulation
through transfusion, transplant, or pregnancy.
Immune hemolytic anemia
Autoimmune Hemolytic Anemia
Production of antibodies that are directed against
the individual’s own RBCs
Four mechanisms:
• Drug adsorption (Hapten)
• Immune complexes
• Membrane modification/Nonspecific protein adsorption
• Autoantibody formation
Drug-Adsorption (Hapten) Mechanism
• Quinidine
• Phenacetin
Membrane Modification
• Cephalosporins
▫ Cephalothin (Keflin)
Autoantibody Formation
• Methyldopa alters the function of T-suppressor cells and
suggest that this upset in the immune system would
allow production of antibody against self.
• Annually:
▫ Mercury thermometers
• Quarterly:
▫ Cell washers
▫ Blood warmers
▫ Centrifuge
• Monthly:
▫ Refrigerated Centrifuge
▫ Alarm activation (ref and freezers)
• Every four hours:
▫ Platelet incubators
Quality Assurance
• Set of planned actions that ensure that systems and
elements that influence the quality of the product or
service are working as expected, individually and
collectively.
Process
(How it happens)
Procedure
(How to do it)
Plan
Improve Implement
Assess
Laboratory Information Systems
Information processing
The proper management of information related to blood
donors, blood components, and patients receiving
transfusions is crucial to ensuring the safety and
traceability of blood products.
Common Information System Acronyms
ACRONYM DEFINITION
• CPU Central processing unit
• HIS Hospital information system
• LIS Laboratory information system
• PC Personal computer
• RAM Random access memory
• ROM Read-only memory
• SOP Standard operating procedure
System Components
Computer system includes three major
components:
• Hardware
• Software
• People
Hardware
Hardware
Three main functions performed by hardware
components:
• Processing
• Input and output
• Storage
Processing Hardware
• Central processing unit
▫ the core of the machine
• Read-only memory (ROM)
▫ contains the “start-up” instructions for the
computer.
• Random access memory (RAM)
▫ an array of chips in which data are temporarily
entered while they are being processed.
Input and Output Devices
Input devices:
▫ Keyboards
▫ Pointing devices
▫ Bar-code readers
▫ Testing instruments
Output devices:
▫ Monitor
▫ Printer
Combined input and output device:
▫ Modem
Software
Tells the computer what to do with all of the information it has
received.
• Operating system software
▫ Controls the computer’s hardware, manipulates the application
software, and coordinates the flow of data to and from disks and
memory.
• Application software
▫ Allows users to perform tasks that are specific to blood bank
operations.
• Interface Software
▫ Allow data to flow between a hospital information system
(HIS) and the blood bank system or between the LIS and the
blood bank.
People
• Human components of a blood bank information system
are the users and at least one person designated as the
system manager.