Review Notes on Blood Banking

BLOOD BANKING
Notes Compiled by: Renz Louie Galanto

Notes Prepared by: Micah Vell Diaz, RMT
REVIEW NOTES ON BLOOD BANKING
OUTLINE
• Historical Perspective • Centrifugation

• Basic Genetics • Effect of Antigen – Antibody Ratio
o Genetics • Dosage
o Human Chromosome • Effect of pH
o Gene • Effect of Temperature
o Locus • Effect of Immunoglobulin Type
o Allele ▪ Enhancement Media / Potentiators
o Polymorphic • Protein Media
o Antithetical • Low Ionic Strength Solution Media (LISS)
o Genotype
• Enzymes
o Phenotype
• Anti-Human Globulin (AHG)
o Dominant
▪ Chemical Reduction of IgG and IgM Molecules
o Recessive
• Dithiothreitol (DTT) and 2-Mercaptoethanol (2-
o Codominant
ME)
o Amorph
o Mendelian’s Law of Inheritance • ZZAP
▪ Law of Dominance ▪ Antihuman Globulin
▪ Law of Independent Segregation • Principles
▪ Law of Independent Assortment • 2 Variations of AHG Test
o Inheritance Pattern ▪ Factors Affecting Antiglobulin Test
o Homozygous • Ratio of Serum to Cells
o Heterozygous • Reaction Medium
o Dosage Effect • Temperature
• Fundamental Concepts • Incubation Time
o The Donation Process • Washing of RBCs
o RBC Biology • Saline for Washing
▪ Normal Chemical Composition and Structure of the • Addition of AHG
RBC membrane • Centrifugation for Reading
▪ Hemoglobin Structure and Function ▪ False Results in AHG
▪ RBC Metabolic Pathways • ABO Blood Group System
• Embden Meyerhof Pathway o The ABO Alle Theories: 3 Allele Theory by Bernstein
• Pentose Phosphate Pathways/Hexose o 4 Allele Theory by Thompson
Monophosphate Pathway o ABO Antigens and Antibodies
• Leubering Rapoport Shunt ▪ ABO Antigens
• Methemoglobin Reductase Pathway • Formation of ABO Antigens
o RBC Preservation ▪ ABO Antibodies
▪ Temperature Requirement o ABO Grouping
▪ Storage Lesion (Loss of RBC viability) ▪ Forward / Direct / Cell Typing
▪ Blood Preservative Contents • Lectins
• Citrate ▪ Reverse / Indirect / Serum Typing
• Adenine ▪ O Group Antibodies
• Glucose o Bombay Phenotype (Oh)
• Phosphate o ABO Subgroups
▪ Approved Preservative Solutions ▪ Subgroups of A
• Additive Solutions ▪ Subgroups of B
• Rejuvenation Solutions o ABO Discrepancies
• Red Cell Freezing ▪ Group I Discrepancies
• RBC Substitutes • Resolution
• Blood Pharming ▪ Group II Discrepancies
o Basic Concepts in Blood Banking • Resolution
▪ Antigen ▪ Group III Discrepancies
▪ Immunogen • Resolution
▪ Antibody
▪ Group IV Discrepancies
• Naturally Occurring
• Categories
• Immune Antibodies
• Resolution
▪ Positive Sign of Antigen – Antibody Reaction in Blood
Banking • RH Blood Group System
▪ Grading of Agglutination Reactions o Nomenclature of the Rh System
▪ Factors that Influence Agglutination Reaction ▪ Fisher – Race (DCE)
• Gene Frequency of Rh Antigen
OUTLINE
• Gene Frequency of Rh Antigen • Temporary Deferral
▪ Wiener (Rh – Hr) • Indefinite Deferral
▪ Rosenfield (Alpha – numeric) • Permanent Deferral
▪ International Society of Blood Transfusion (ISBT) o Blood Collection
o Rh Typing ▪ Collection of Whole Blood
o Weak D Typing ▪ Submission of Whole Blood to Blood Bank/Center
o Causes of Weak - D ▪ Labelling of Blood Bags
▪ C trans to RHD ▪ Donor Blood Unit Processing
▪ Del (D -) • Blood Component Preparation
▪ Rh Null (--/--) o Separation of Component
▪ Rh mod o Open System and Close System
▪ F(ce)
o Components
▪ Genetic Weak D
▪ Partial D (D mosaic)
o Summary
o LW Blood Group System
o Transfusion
▪ Hemapheresis / Apheresis
▪ Anti – LW reaction
• Methods
• Strong
• Frequency of Donation
• Weak
• Transfusion Reaction
• Negative
• Transfusion – Transmitted Diseases
• Major Blood Groups
o Lewis Blood Group System (ISBT 007) • Blood Banking Techniques
▪ Rules Regarding Le antigen Expression o Antibody Screening
o MNS Blood Group System (ISBT 002) o Antibody Identification / Panel Testing
▪ MN Antigens o Crossmatching
▪ Ss Antigens ▪ Major Crossmatching
▪ MNS Antibodies ▪ Minor Crossmatching
o P Blood Group System (ISBT 003) ▪ Types of Crossnatch
▪ P1 Antigen o Serologic Techniques
▪ P Antibodies o Technologies in Blood Banking
▪ Disease Associations ▪ Tube Testing
o I Blood Group System (ISBT 027) ▪ Gel Technology
▪ I Antigen ▪ Solid Phase Technology
▪ I Antibodies ▪ Affinity Column Technology
o Kell Blood Group System (ISBT 006) • Hemolytic Disease of the Fetus and Newborn
▪ Kell Antigens o Diagnosis and Treatment of HDFN
▪ Kell Antibodies o Etiology
o Duffy Blood Group System (ISBT 008) o Rh HDFN
▪ Duffy Antigens ▪ During Gestation and Delivery
▪ Duffy Antibodies ▪ Factors Affecting Immunization and Severity of HDFN
o Kidd Blood Group System (ISBT 009) o Pathogenesis
▪ Jka and Jkb Antigens ▪ Hemolysis, Anemia, and Erythropoiesis
▪ Kidd Antibodies (anti Jka and Jkb) ▪ Bilirubin
o Lutheran Blood Group System (ISBT 010) o Diagnosis and Management
• Minor Blood Groups ▪ Serologic Testing of the Mother
o Summary ▪ Color Doppler Middle Cerebral Artery Peak Systolic
• Blood Donor and Selection Processing Velocity
o The Donation Process ▪ Cordocentesis
▪ Registration ▪ Amniocentesis
▪ Interview and Appearance ▪ Intrauterine Transfusion
o Requirements for Allogeneic Donation ▪ Phototherapy
o Adjustment of Blood Volume and Blood Anticoagulant to ▪ Intravenous Immune Globulin
donors weighing <110 pounds ▪ Exchange Transfusion
▪ Serologic Testing for the Newborn Infant
o Autologous Transfusion
▪ Newborn Transfusions
o Methods of Hemoglobin Determination
o Mechanism of Action of RhIg
o Types of Blood Donation o ABO – HDN
o Types of Autologous Donation o Comparison of ABO Vs Rh HDFN
o Deferral Guidelines for Allogeneic Donation • Autoimmune Hemolytic Anemias (AIHA)
▪ Types of Deferral o Warm AIHA and Cold AIHA
HISTORICAL PERSPECTIVE POLYMORPHIC______________________
● Having two or more possible allele at a locus
1492 – Pope Innocent VII Recipient of the first blood transfusion in
history ANTITHETICAL_____________________
1840 – James Blundell Performed the first successful blood ● Opposite antigens encoded at the same locus
transfusion to patient
1869 – Braxton Hicks Recommended sodium phosphate as
anticoagulant
GENOTYPE__________________________
● Total genetic composition of an individual derived from maternal
1901 – Karl Landsteiner Discovered the ABO Blood Groups and paternal genes
1902 – Von Descatello and Discovered the fourth blood group (Type AB) ● Actual genetic make up
Sturli
1913 – Edward Used vein-to-vein transfusion by using
Lindemann multiple syringes and a special cannula PHENOTYPE________________________
1913 – Lester Unger Designed syringe – valve apparatus; ● Detectable or expressed characteristics of genes
unassisted transfusion ● Actual expression of the gene
1914 – Albert Hustlin Used sodium citrate as an anticoagulant
solution for transfusions DOMINANT__________________________
1915 – Richard Lewison Determined the minimum amount of citrate ● Only one allele must be inherited for it to be expressed
needed for anticoagulant ● Gene product always present
1916 – Rous and Turner Introduced a citrate – dextrose solution for the
preservation of blood
1940 – Karl Landsteiner Discovered Rh blood group
RECESSIVE__________________________
● Expressed only in homozygous state, not expressed in the presence
and Alex Wiener
of a dominant gene
1941 – Dr. Charles Drew Appointed director of the first American Red
Cross Blood Bank at Presbyterian Hospital
1943 – Loutit and Introduced the formula for the preservative CODOMINANT_______________________
Mollison acid-citrate-dextrose (ACD) ● Term used to describe a pair of genes in which neither is dominant
1945 – Robin Coombs, Describe the use of Anti-Human Globulin over the other
Arthur Mourant, and Rob (AHG) ● Both genes expressed equally
Race
1950 – Carl Walter and
William Murphy
Introduced the use of flexible sealed plastic
bag for blood transfusion
AMORPH____________________________
● Gene with no observable manifestation or trait
1957 – Gibson Introduced citrate phosphate – dextrose
(CPD)
1972 – Herb Cullis Invented apheresis machine MENDELIAN’S LAW OF INHERITANCE__
1975 – George Kohler and Discovered monoclonal antibodies thru
Cesar Milstein Hybridoma technology LAW OF DOMINANCE
● States that hybrid offspring will only inherit the dominant trait in the
phenotype
BASIC GENETICS ● The alleles that are suppressed are called the recessive traits whole
the alleles that determine the trait are known as dominant traits
GENETICS___________________________
● Study of transmission of inherited characteristics LAW OF INDEPENDENT SEGREGATION
● Study of inheritance ● States that during the production of gametes, two copies of each
hereditary factor segregate so that offspring acquire one factor from
HUMAN CHROMOSOME_____________ each parent
● 46 in each nucleated cells (22 pairs of autosomes and 1 set sex
chromosome) LAW OF INDEPENDENT ASSORTMENT
o Autosomal genes: genes expressed with equal frequency in ● States that traits inherited from different chromosomes expressed
males and females separately and discretely
o Sex – linked genes: genes carried on the X chromosome
INHERITANCE PATTERN_____________
GENE_______________________________ ● Predicted using Punnett Square
● Basic unit of inheritance
A segment of DNA arranged along the chromosome at a specific
●
position called locus
HOMOZYGOUS______________________
● Individuals inherit identical alleles at the same locus
LOCUS______________________________ HETEROZYGOUS____________________
● Position of a gene in a chromosome
● Individuals inherit different alleles at the same gene locus
ALLELE_____________________________ DOSAGE EFFECT____________________

● One of two or more alternative genes, may be present at a given
locus in a chromosome ● Agglutination reactions are stronger for homozygous cells and
● Alternate forms of a gene slightly weaker for heterozygous cells
o Rh, Kidd, Duffy, MNS, Lutheran
PENTOSE PHOSPHATE PATHWAYS / HEXOSE
FUNDAMENTAL CONCEPTS MONOPHOSPHATE PAHTWAY
● Traditional whole blood collection volume per unit: 450 mL +/- 10%
of blood (1 pint) ● Provides 10% of ATP and reduce glutathione
● Current whole blood collection volume per unit: 500 mL +/10% of
blood (modified plastic collection systems) LEUBERING RAPOPORT SHUNT
● Sizes of blood bag:
o 350 mL = 49 mL anticoagulant - preservative ● Permits accumulation of 2,3 – DPG; off-loads oxygen to tissue
o 450 mL = 63 mL anticoagulant - preservative
o 500 mL = 70 mL anticoagulant - preservative METHEMOGLOBIN REDUCTASE PATHWAY
● Needle size/length: 16 gauge / 1 – 1.5 – inch needle
● AC: Blood ratio: 1 part AC: 7 parts blood ● Maintains heme iron of the hemoglobin in ferrous state
● Maximum allowable amount of blood to be extracted: 10.5 mL/kg
donor’s weight RBC PRESERVATION_________________
● For a 110-pound donor, a maximum of 525 mL can be collected, ● Goal: Maintain RBC viability during storage
including samples drawn for processing ● FDA requires an average of 24-hour post transfusion RBC survival
● Donation average length of time: 8 – 15 minutes of more than 75%
● Total blood volume of most adults is 10 – 12 pints ● 70% of red cells must remain viable at the end of the allowed storage
● 1 pint of blood can be replenished its volume within 24 hours time
● Period of replacement of the collected RBCs: 1 – 2 months
● Donors can donate whole blood every 8 weeks TEMPERATURE REQUIREMENT
● REMEMBER: Donated Blood is FREE, but fee is charged for each
unit to cover the costs associated with collecting, storing, testing, ● 1 – 6 oC – storage temperature of liquid whole blood
and transfusing blood ● 1 – 10oC – shipping/transport temperature of liquid whole blood
STORAGE LESION (LOSS OF RBC VIABILITY)

THE DONATION PROCESS____________
● 1. Educational Materials – contains information on the risks of ● Increased: Hemoglobin, Lactic acid, Potassium (HeLP)
infectious diseases transmitted by blood transfusions, including the ● Decreased: pH. Glucose, ATP, 2,3 – DPG, Sodium
symptoms and signs of AIDs, is given to each prospective donor to
read BLOOD PRESERVATIVE CONTENTS
● 2. The Donor Health History Questionnaire – A uniform donor
history questionnaire, designed to ask questions that protect the CITRATE
health of both the donor and the recipient, is given to every donor
● 3. The Abbreviated Physical Examination – includes blood pressure, ● Binds calcium
pulse, and temperature readings; hemoglobin or hematocrit level; ● Serves as anticoagulant
and the inspection of the arms for skin lesions.
ADENINE
RBC BIOLOGY_______________________
● Increase ADP levels
NORMAL CHEMICAL COMPOSITION AND STRUCTURE OF ● For ATP synthesis
THE RBC MEMBRANE
GLUCOSE
● 52% protein; 40% lipid; 8% carbohydrates
● Semipermeable lipid bilayer ● Serves as food for the cells
● Phospholipid – main membrane content
● Integral protein – extend from the outer surface; span the entire PHOSPHATE
membrane to the inner cytoplasmic side of the RBC (Glycophorins
– GPA, GPB, GPC) ● Source of 2,3 – DPG
● Peripheral protein – located and limited to the cytoplasmic surface
of the membrane forming the RBC cytoskeleton (Spectrin, Actin, APPROVED PRESERVATIVE SOLUTIONS
Ankyrin)
Preservative Abbrev. Shelf-life Usage
HEMOGLOBIN STRUCTURE AND FUNCTION Acid-Citrate-Dextrose ACD 21 Apheresis
Citrate-Phosphate-Dextrose CPD 21 Whole Blood
● Structure: 2 pairs of globin chains; 4 heme groups Citrate-Phosphate-Double CP2D 21 WB
● Normal types of adult hemoglobin: Dextrose
o 92 – 95% HbA = 2 alpha, 2 beta chains Citrate-Phosphate-Dextrose- (CPDA-1) 35 WB
o 2 – 3% HbA2 = 2 alpha, 2 delta chains Adenine
o 1 – 2 % HbF = 2 alpha, 2 gamma chains *Heparin – used for by-pass surgery; 2 days shelf-life
● Function: Gas transport
● Carries oxygen from the lungs to the peripheral tissue ADDITIVE SOLUTIONS
● Returns carbon dioxide from the tissue to the lungs for excretion
● Support red cell survival up to 42 days
RBC METABOLIC PATHWAYS ● Must be added to RBCs within the first 72 hours of storage
● Added after removal of plasma with/without platelet
EMBDEN MEYERHOF PATHWAY ● 100 mL additive solution is used for a 450 mL blood bag
● Final hematocrit: 55 – 65%; (hct w/o additive: 65 – 80%)
● Anaerobic glycolysis that provides 90% of ATP needed by RBCs ● Content: Saline, Adenine, Glucose, Mannitol (SAGM); or Saline,
Adenine, Glucose, Citrate (SAGC)
ADDITIVE ABBREV. STORAGE NATURALLY OCCURRING
Adsol (Baxter Healthcare) AS-1 (SAGM) 42
Nutricel (Pall corporation) AS-3 (SAGC) 42 ● Found in serum of individuals without being exposed to matching
Optisol (Terumo corporation) AS-5 (SAGM) 42 antigen
● Ex.: ABO, Ii, Lewis, P, MN Antibodies
REJUVENATION SOLUTIONS
IMMUNE ANTIBODIES
● Used to restore levels of ATP and 2,3-DPG, levels in RBCs stored
in CPD, CPDA-1, AS-1 ● Found in serum of individuals after being exposed to matching
● Solution can be used up to 3 days after expiration of red cells antigen
(rejuvenation for 1-4 hours at 37oC) ● Ex.: Rh, Kell, Kidd, Duffy, Ss antibodies
● REJUVESOL: only FDA-approved rejuvenation solution in US
● Content: Phosphate-Inosine-Glucose-Pyruvate-Adenine (PIGPA) POSITIVE SIGN OF ANTIGEN – ANTIBODY REACTION IN
Phosphate-Inosine-Pyruvate-Adenine (PIPA) BLOOD BANKING
● Wash rejuvenated RBCs to remove inosine ● 1. Hemagglutination – most common
● Freeze rejuvenated RBCs: shelf-life for 10 years using CPD and ● 2. Hemolysis
CPDA-1; shelf-life if 3 years using AS-1
GRADING OF AGGLUTINATION REACTIONS
RED CELL FREEZING
GRADE TUBE METHOD GEL TEST METHOD
● Used for autologous units and storage of rare blood cell types 4+ Orie solid Solid band of agglutinated RBCs at the
● Storage: 10 years (-65oC) agglutinate, clear TOP of the gel column, no RBCs are
● Requirement: addition of cryoprecipitate agent (glycerol) to RBCs supernatant visible at the bottom
that are less than 6 days old 3+ Several large Predominant amount of agglutinated
● Deglycerolization – done prior to transfusion to avoid infusing the agglutinates, clear RBCs NEAR THE TOP/UPPER HALF of
hypertonic glycerol supernatant the gel column, few agglutinates below
● Red cell washing with decreasing concentration of saline: 12% -> 2+ Medium-sized RBC agglutinates that are dispensed
1.6% -> 0.85 (NSS) + 0.2% dextrose agglutinates, clear THROUGHOUT the gel column, few
ADVANTAGE HIGH GLYCEROL LOW GLYCEROL background agglutinates at the bottom
Initial freezing -80 oC -196 oC 1+ Small RBC agglutinates that are predominantly
temperature agglutinates, in the LOWER HALF of the gel column,
Control the freezing No YES turbid background some RBCs at the bottom
rate 0 No agglutination RBCs form a well-delineated pellet at the
Type of freezer Mechanical Liquid Nitrogen bottom of the microtube. The gel above the
Maximum storage -65 oC -120 oC RBC pellet is clear
temperature
Shipping Dry Ice Liquid Nitrogen
requirements
RBC SUBSTITUTES
● Carry oxygen in the absence of RBCs

● Hemoglobin – based oxygen carrier
● Perfluorocarbons (PFCs) – hydrogen atoms are replaced by
FLUORINE (0.2 um in dm)
BLOOD PHARMING
● Arteriocyte – O-negative prod

● NANEX – turn hematopoietic stem cells from umbilical cord into
O-type, Rh negative RBC
BASIC CONCEPTS IN BLOOD BANKING

ANTIGEN
● “Antibody Generator”
● Recognized as foreign by the body, capable or incapable of inducing
immune response
IMMUNOGEN
● Antigens which are always capable of immune response FACTORS THAT INFLUENCE AGGLUTINATION REACTIONS
ANTIBODY
CENTRIFUGATION
● Protein
● Capable of neutralizing viruses, binding bacterial cell, and any ● Simplest and most common technique to enhance agglutination
foreign or waste of the body
EFFECT OF ANTIGEN – ANTIBODY RATIO o Polyclonal – pool of heterogenous anti – IgG from many rabbits
o Monoclonal – pool of anti-IgG from a single clone of plasma
● Prozone: Antibody excess; remedy: SERUM DILUTION cells
● Postzone: Antigen excess; remedy: INCREASE SERUM-TO-CELL ▪ Prepared by Hybridoma Technology
RATIO ▪ Uses mice
DOSAGE CHEMICAL REDUCTION OF IGG AND IGM MOLECULES
● Heterozygous alleles has less antigens on red cells (weak

agglutination) DITHIOTHREITOL (DTT) AND 2 – MERCAPTOETHANOL (2-
● Homozygous alleles has more antigens on red cells (stronger ME)
agglutination
o Demonstrates dosage effect: Duffy, M, N, S, Kidd, Rh, ● Thiol – reducing agents that break the disulfide bonds of the J
Lutheran (joining) chain of the IgM molecule but leave the IgG molecule
intact
EFFECT OF PH
ZZAP
● Ideal pH: 6.5 – 7.5 ● Thiol reagent plus a proteolytic enzyme
● Causes the dissociation of IgG molecules from the surface of
EFFECT OF TEMPERATURE sensitized RBCs and alters the surface antigens of the RBC
● IgM reacts best at cold temperature (4-22oC) ANTIHUMAN GLOBULIN

● IgG reacts best at warm temperature (37oC)
● Also referred to as Coomb’s Test
EFFECT OF IMMUNOGLOBULIN TYPE
● IgM = agglutinates red cells even in 0.85% NSS

● IgG = requires enhancement media potentiators
ENHANCEMENT MEDIA / POTENTIATORS
PROTEIN MEDIA
● Increases the dielectric constant, which reduces the zeta potential

o 22% Albumin
▪ Causes agglutination by adjusting zeta potential between
red cells
▪ Incubated at 37oC for 15 – 30 minutes
o Polyethylene glycol (PEG)
▪ Reduces false – positive reactions
▪ More effective than albumin
▪ Incubated at 37oC for 10 – 30 minutes
o Others: Polybrene, Polyvinylpyrrolidone (PVP), Protamine
LOW IONIC STRENGTH SOLUTION MEDIA (LISS)
● Decreases the ionic strength of a reaction medium and so reduce the

zeta potential
● Increase rate of antibody uptake during sensitization
● Incubated at 37oC for 5 – 15 minutes
PRINCIPLES
ENZYMES
● Antibody molecules and complement components are globulins
● ENHANCES antibody reactivity to antigens: Rh, Kidd, P1, Lewis, ● Injecting an animal with human globulin stimulates the animal to
and I antigen produce antibody to foreign protein
● DESTROYS reactivity to red cell antigens: Fya, Fyb, M, N, S o Serologic tests employ a variety of AHG reagents reactive with
antigens various human globulins, including anti – IgG antibody to the
● Ex.: Ficin (fig plant), Trypsin (pig stomach), Bromelin (pineapple), C3d component of human complement, and polyspecific
Papain (papaya) reagents that contain both anti – IgG and anti – IgG and anti –
C3d activity
ANTI-HUMAN GLOBULIN (AHG)
2 VARIATIONS OF AHG TEST
● Designed to determine if RBCs are coated with antibody or
complement or both ● 1. Direct Antiglobulin Test (DAT)
o Polyspecific AHG – can determine if RBCs have been o Detects in vivo sensitization of RBCs with IgG or complement
sensitized with IgG antibody or complement (components C3b components
or C3d) or both o Detects clinical conditions: Hemolytic disease of the newborn
o Monospecific AHG – react only with RBCs sensitized with IgG (HDN), Hemolytic transfusion reaction (HTR), Autoimmune
or complement and drug-induced hemolytic anemia (AIHA)
2. Indirect Antiglobulin Test (IAT)
●
o Determine in vitro sensitization of RBCs and is used in the
THE ABO ALLE THEORIES: 3 ALLELE
following situations: THEORY BY BERNSTEIN_____________
▪ Antibody detection (Compatibility test and Antibody Phenotype Possible Genotypes
screening test) A AA, AO
▪ Antibody identification (Antibody panel test) B BB, BO
▪ Antibody titration, red cell phenotype AB AB
O OO
FACTORS AFFECTING ANTIGLOBULIN TEST
4 ALLELE THEORY BY THOMPSON____

RATIO OF SERUM TO CELLS
Phenotype Possible Genotypes
A1 A1 A1, A1O, A1A2
● 40:1 (drops of serum and 1 drop of a 5% volume of solute per
volume of solution (v/v) suspension of cells) A2 A2 A1, A2O
A1B A1B
REACTION MEDIUM A2B A2B
B BB, BO
● Albumin: allow Ab-coated cells to come contact with each other O OO
● LISS: enhance Ab update; shorter incubation time Alleles: A1, A2, B, O
● PEG: removes water thereby concentrating Ab
ABO ANTIGENS AND ANTIBODIES____
TEMPERATURE
ABO ANTIGENS
● IgG optimal at 37oC
● Found on RBCs, lymphocytes, platelets, endothelial cells, and
INCUBATION TIME epithelial cells
● Developed at 37th day of fetal life, full expression occurs between 2
● Saline: 30 – 120 minutes – 4 years of age
● 22% Albumin: 15 – 60 minutes ● ABH genes code for the production of glycosyl transferase catalyzes
● LISS/PEG: 10 – 15 minutes the addition of specific immunodominant carbohydrate to the type 2
chain
WASHING OF RBCS ● Precursor Oligosaccharide Chains:
Type 1 Type 2
● Wash 3 times before addition of AHG Linkage Beta-1,3 Beta-1,4
Origin Plasma Seen on erythrocytic precursors
SALINE FOR WASHING Controlling genes H, A, B, Se, and Lewis H, A, and B
● Fresh and have a pH of 7.2 – 7.4 FORMATION OF ABO ANTIGENS
ADDITION OF AHG Gene Glycosyltransferase Immunodominant Acceptor Antigen

Sugar
● Added immediately after washing H a-2-L- L – fucose Precursor H
fucosytransferase
CENTRIFUGATION FOR READING A a-3-N- N-acetyl-D- H A
acetylgalactos galactosamine
● 1000 RCF for 20 seconds aminyltransferase
B a-3-D-galactosyl D-galactose H B
FALSE RESULTS IN AHG transferase
FALSE POSITIVE FALSE NEGATIVE AB a-3-N- N-acetyl-D- H AB
Clotted specimen Inadequate washing of cells acetylgalactos galactosamine
Bacterial contamination of cells Inadequate incubation conditions aminyltransferase
Metal contamination of saline Old serum samples a-3-D-galactosyl D-galactose
Dirty glassware Serum/AHG not added transferase
Overcentrifugation Undercentrifugation 0 ________________ ---------------- -------- Unaltered
H antigen
Overreading Cells suspension too weak or too heavy
Contaminated AHG Non-reactive AHG ● H gene – FUT1, located in chromosome 19
● Four H antigen subtypes: H1, H2, H3, H4
ABO BLOOD GROUP SYSTEM ● Amount of H antigen: O > A2 > B > A2B > A1 > A1B
● Secretor Gene (Se gene) – FUT2, located at chromosome 19, codes
● ISBT 001 for the production of a-2-L-fucosyltransferase which is expressed in
● First discovered and most important of all blood groups in tissues related to exocrine secretions
transfusion practice ● Fluids in which ABH substances can be detected: Saliva, tears, milk,
● Only blood group system in which individuals have antibodies in digestive juices, bile urine, amniotic fluid, pathologic fluid
their serum to antigens that are absent from their RBCs ● RBC vs. Secretory ABH Antigens
● Causes the most severe HTR
● Frequency of ABO phenotypes O > A > B > AB ABH on RBC ABH on secretion
● The three genes that code for A, B, O are located at the long arms of Type 2 precursor Type 1 precursor
chromosome 9
Glycolipids, glycoproteins, or Glycoprotein 3+ Several large aggregates, clear background
glycosphingolipids 2+ Small to medium-sized aggregates, clear background
Enzyme: a-2-L- Enzyme: a-2-L- 1+ Small aggregates, turbid reddish background
fucosyltransferase by H gene fucosyltransferase by Se gene W+ Tiny aggregates, turbid reddish background
(FUT1) (FUT2) M+ Mixed field – Any degree of agglutination in a sea of
unagglutinated cells
ABO ANTIBODIES HEM Hemolysis
● Naturally occurring and predominantly IgM in nature
● Pre-formed and developed between 3 – 6 months of age; peaks REVERSE / INDIRECT / SERUM TYPING
between 5 – 10 years of age and declines later in life ● Use known A1 and B antigens and testing the serum of the patients
● Immunogenic Ab: Anti A,B, Anti – A1 FOR DETECTING AB
Blood Group Antibody Produced Characteristics O GROUP ANTIBODIES

A Anti-B IgM
B Anti - A Naturally occurring ● Reagent: Known A and B cells; human source; 2 – 5% red cell
antibodies suspension
React at room temperature ● Specimen: Serum
CANNOT cross the Blood Group Antibody A1 cells B cells Interpretation
placenta A Anti-B - + A
AB --------- B Anti-A + - B
O Anti – A Mostly IgM AB NONE - - AB
Anti – B Mostly IgM O Anti-A, + + O
Anti – A, B Newly found antibody; Anti-B
predominantly IgG; can
cause ABO HDFN BOMBAY PHENOTYPE (Oh)___________
● hh genotype
ABO GROUPING_____________________ ● No H antigens formed = No A nor B antigens
● Principle: Hemagglutination ● Phenotypes as blood group O
● Antibodies present: Anti – A, Anti – B, Anti – H
FORWARD / DIRECT / CELL TYPING ● Can only be transfused with blood from another Bombay (Oh)
Phenotype Forward Typing Reverse Typing
● Use known sources of antisera to DETECT ANTIGENS of Anti-A Anti-B Anti-H A cells B cells O cells
individual’s red cells O - - + + + -
● Reagent: Typing sera (Anti – A, anti – B) Oh - - - + + +
● Specimen: Washed Red Cells
o Anti – A Reagent: Monoclonal antibody; IgM; CLEAR BLUE
color; Blue dye: Bromphenol blue, Thymol blue, Patent blue ABO SUBGROUPS____________________
o Anti – B Reagent: Monoclonal antibody; IgM; CLEAR ● Differ in the amount of antigen present on the RBC membrane as a
YELLOW color; Yellow Dye: Acriflavine, Tartrazine yellow result of less effective enzymes
Blood Antigen Anti - Anti - Anti – Interpretation
Group A B A, B SUBGROUPS OF A
A A + - + A
Phenotype Antigens Population Reaction with
B B - + + B
present frequency Anti-A (from B Anti-A1
AB AB + + + AB
sera) lectin
O NONE - - - O
A1 A1 A2 80% + +
A2 A 20% + -
LECTINS
● 1 – 8% of A2, 25% of A2B, can form Anti – A1
● Other subgroups of A
● Proteins present in plants, which bind specifically to carbohydrate
determinants and agglutinate RBCs through their cell surface of Phenotypes Description
oligosaccharide determinants A3 Mixed field agglutination with anti-A and/or anti-A,B
o Lectins for ABO antigens: Ax Weak agglutination with anti-A, B only
▪ Ulex europaeus – Anti – H Aend <10% red cells show very weak mf agglutination
▪ Dolichus biflorus – Anti – A Am No agglutination with anti-A and anti-A, B
▪ Griffonia simplicifolia (Bandeiraea simplicifolia) – Anti Secretors demonstrate quantities of A substance in saliva
–B Ay No agglutination with anti-A and anti-A, B
o Lectins for antigen causing polyagglutination: Secretors contains small amount of A substance in saliva
▪ Arachis hypogaea – Anti – K, Anti – Tk Ael No agglutination with anti-A and anti-A, B
▪ Glycine soja (Glycine max) – Anti – T, Anti – Tn, Anti – Secretors contains only H substance and no A substance in saliva
Cad
▪ Salvia sclarea – Anti – Tn SUBGROUPS OF B
▪ Salvia horminum – Anti – Tn, Anti – Cad Phenotypes Description
o Lectins for other blood group antigens:
B3 Mixed field agglutination with anti-B and/or anti-A,B
▪ Iberis amara – Anti – M
Bx Agglutination with anti-A, B (wk/0 with anti-B)
▪ Vicia graminea and Bauhinia species – Anti – N
Bm No agglutination with anti-B and anti-A, B
Grade Definition
Secretors demonstrate quantities of B substance in saliva
Macroscopic
Bel No agglutination with anti-B and anti-A, B
4+ One solid aggregate or clump of cells
Secretors contains only H substance and no B substance ● Use enzyme treated RBCs
in saliva
GROUP III DISCREPANCIES
ABO DISCREPANCIES________________ ● Protein or Plasma Abnormalities Resulting to Rouleaux Formation
● Common sources of Technical Errors resulting to Discrepancies o Elevated levels of globulin from certain disease states (ex.
o Incorrect or inadequate identification of blood specimens, test Multiple Myeloma, Waldenstrom’s macroglobulinemia)
tubes, or slides o Elevated levels of fibrinogen
o Cell suspension either too heavy or too light o Plasma expanders
o Clerical errors or incorrect recording of results o Wharton’s Jelly
o A mix-up in samples
o Missed observation of hemolysis RESOLUTION
o Failure to add reagents
o Failure to add sample ● Wash RBCs with saline several times (at least 6 – 8 times)
o Failure to follow manufacturer’s instruction
o Uncalibrated centrifuge GROUP IV DISCREPANCIES
o Overcentrifugation or under centrifugation ● Miscellaneous
o Contaminated reagents o Polyagglutination – refers to agglutination of altered RBCs by
o Warming during centrifugation a large proportion of ABO-compatible adult human sera
● RESOLUTION ▪ RBCs show agglutination with most adult sera but no
o If the initial test was performed using RBCs suspended in agglutination with cord sera
serum or plasma, repeat testing the same sample using a saline
suspension of RBCs CATEGORIES
o Acquire information regarding the patient’s age, diagnosis,
transfusion history, medications, and history of pregnancy ● Acquired
o New sample should be draw if discrepancy persists o Microbially associated: passive adsorption of bacterial
products: T, Th, Tk, Tx, Acquired B, VA
GROUP I DISCREPANCIES o Non-microbially associated: mutation of hematopoietic tissueL
● Weakly reacting or Missing Antibodies Tn
o Newborns ● Inherited – Cad, NOR, Hemoglobin M-Hyde Park, Hereditary
o Elderly patients Erythroblastic Multinuclearity with Positive Acidified Serum
o Patients with a leukemia (e.g. chronic lymphocytic leukemia) (HEMPAS)
or lymphoma (e.g. malignant lymphoma) o Cold reactive antibodies
o Patients using immunosuppresive drugs o Unexpected ABO isoagglutinins
o Patients with congenital or acquired agammaglobulinemia or o Antibodies other than anti – A and anti – B may react to from
immunodeficiency diseases antigen – antibody complexes that may then adsorb into
o Patients with bone marrow or stem cell transplantations patient’s red cell
o Plasma transfusion or exchange transfusion o RBCs with the cis “AB phenotype”
o ABO subgroups
RESOLUTION
RESOLUTION
● RBC (sample) = wash with saline (37 degrees Celsius); 0.01 M
● Obtain the patient’s history Dithiothreitol (DTT)
● Incubate the patient serum with reagent A1 and B cells at room ● Serum (sample) = warm at 37 degrees Celsius then read results at 37
temperature for approximately 15 to 30 minutes or by adding one or degrees Celsius
two drops more plasma or serum to the test
● Incubated at 4oC for 15 minutes (if still no reaction after Table 6-17. Example of ABO Discrepancy Seen with Weak or Missing
centrifugation) Antibodies
FORWARD GROUPING REVERSE GROUPING
GROUP II DISCREPANCIES REACTION OF PATIENT’S REACTION OF PATIENT’S
CELL WITH CELL WITH
● Weakly reacting or Missing Antibodies Anti - A Anti - B A1 cells B cells
o Subgroups of A (or B)
Patient 0 0 0 0
o Leukemias, Hodgkin’s Disease
Patient’s probably group: O (elderly patient or newborn)
o The “acquired B” phenomenon
Note: The absence of agglutination with reagent cells in the reverse type
▪ Occurs when bacterial enzyme modifies
is because the production of ABO antibodies can be weak or absent in the
immunodominant sugar A (N-acetyl-D-galactosamine)
elderly
into D-galatosamine (similae to D-galactose) which cross
Resolution: (1) check the age of the patient, (2) increase incubation time
reacts with anti-B antisera
to 30 minutes (not appropriate for newborn sample), (3) lower
● Resolution: use monoclinal anti-B clone (ES4);
temperature to 4oC for 15 minutes (include O cells and an autocontrol)
treat RBCs using acetic anhydride
Table 6-18. Serologic Reactions Typical of Leukemia
RESOLUTION
FORWARD GROUPING REVERSE GROUPING
● Use 2 – 5% RCS REACTION OF REACTION OF
● Incubate the patient serum with reagent A1 and B cells at room PATIENT’S CELL WITH PATIENT’S CELL WITH
temperature for approximately 15 to 30 minutes or by adding one or Patient Phenotype Anti - A Anti - B A1 cells B cells
two drops more plasma or serum to the test A +mf 0 0 3+
● Incubated at 4oC for 15 minutes (if still no reaction after B 0 +/+ 4+_ 0
centrifugation)
Note: Weak reactivity with anti – A and anti – B is because the disease, Resolution: (1) Test cells with anti-A1 lectin; (2) Test serum with A1, A2
leukemia has resulted in the weakened expression of the corresponding and O cells; (3) Run an autocontrol
antigen
Table 6-25. Example of ABO Discrepancy Caused by Unexpected Non-
Table 6-19. Example of ABO Discrepancy Caused by an Acquired B ABO Alloantibody
Antigen FORWARD GROUPING REVERSE GROUPING
FORWARD GROUPING REVERSE GROUPING REACTION OF PATIENT’S REACTION OF
REACTION OF PATIENT’S REACTION OF CELL WITH PATIENT’S CELL WITH
CELL WITH PATIENT’S CELL WITH Anti - A Anti - B A1 cells B cells
Anti - A Anti - B A1 cells B cells Patient 4+ 4+ 1+ 1+
Patient 4+ 2+ 0 4+ Patient’s probably group: AB
Patient’s probably group: A Note: Reactions with patient serum are due to non-ABO alloantibody
Note: Patient RBCs have acquired a B-like antigen that reacts with agglutinating an antigen other than A1 and B on reagent red cells
reagent anti – B and is associated with cancer of the colon or other Resolution: (1) Run an antibody screen and panel
diseases of the digestive tract
Resolution: (1) Acidify Anti-B reagent to a pH of 6, (2) Run DAT, (3)
Run autocontrol RH BLOOD GROUP SYSTEM
● ISBT 004
Table 6-20. Example of ABO Discrepancy Caused by Low – Incidence ● Rhesus Blood Group System
Antibodies in the Reagent Antisera ● Discovered by Landsteiner and Weiner in 1940
FORWARD GROUPING REVERSE GROUPING ● Genes are located on the short arm of chromosome 1
REACTION OF PATIENT’S REACTION OF ● Primary cause of hemolytic disease of the fetus and newborn
CELL WITH PATIENT’S CELL WITH (HDFN) and a significant cause of hemolytic transfusion reaction
Anti - A Anti - B A1 cells B cells (HTR)
● Rh antigen are non-glycosylated proteins and are integral part of
Patient 0 1+ 4+ 4+
RBC membrane
Patient’s probably group: O ● Immunogenicity: D > c > E > C > e
Note: Reaction with anti-B in the forward type is due to agglutination ● Rh antibodies: IgG (reacts at 37 degrees Celsius); do not bind
between a low-incidence antibody in reagent anti-b and the corresponding complement; EXTRAVASCULAR hemolysis
antigen on the patient’s cells
Resolution: Use a different lot number for reagent anti – B
NOMENCLATURE OF THE RH SYSTEM
Table 6-22. Example of ABO Discrepancy Caused by Rouleaux
Formation FISCHER – RACE (DCE)
FORWARD GROUPING REVERSE GROUPING ● The antigens were produced by 3 closely linked set of alleles, each
REACTION OF REACTION OF gene was responsible for producing a product or antigen on the red
PATIENT’S CELL WITH PATIENT’S CELL WITH cell surface
Anti - A Anti - B A1 cells B cells
Patient 4+ 4+ 2+ 2+ GENE FREQUENCY OF RH ANTIGEN
Patient’s probably group: AB Gene Frequency
Note: Agglutination with A1 and B cells in reverse type is due to D 85%
rouleaux formation as a result of increased serum protein or plasma d or absence of D antigen 15%
abnormalities C 70%
Resolution: (1) Microscopic examination, (2) Saline replacement
E 80%
technique, (3) Wash cells with saline three times, (4) Run antibody
c 30%
screen
e 98%
Patient 2+ 4+ 4+ 2+
Patient’s probably group: B WIENER (RH – HR)
Note: Reaction with anti-A in forward type is due spontaneous
agglutination of antibody coated cells; reaction with B cells in reverse ● The gene responsible for defining Rh actually produced an
type is due to cold autoantibody (e.g. anti – I) reacting with I antigen on agglutinogen that contained a series of blood factor, in which each
B cells factor is an antigen recognized by an antibody
Resolution: (1) Wash patient cells with warm saline and retest, (2) Run Gene Agglutinogen Shorthand Blood Factors Fisher - Race
DAT and autocontrol, (3) Run antibody screen Rh0 Rh0 R0 Rh0hr’hr” Dce
1
Rh Rh1 R1 Rh0rh’hr” DCe
Table 6-24. Example of ABO Discrepancy Caused by an Unexpected Rh2 Rh2 R2 Rh0hr’rh” DcE
ABO Isoagglutinin Rhz Rhz RZ Rh0rh’rh” DCE
FORWARD GROUPING REVERSE GROUPING rh rh r hr'hr” dce
REACTION OF PATIENT’S REACTION OF rh' rh' r' rh'hr” dCe
CELL WITH PATIENT’S CELL WITH rh" rh" r" hr'rh” dcE
Anti - A Anti - B A1 cells B cells rhy rhy ry rh'rh” dCE
Patient 4+ 4+ 1+ 0
ROSENFIELD (ALPHA – NUMERIC)
Patient’s probably group: A2B
Note: Reactions with patient serum are due to anti-A, agglutinating A1 ● Has no genetic basis
reagent red cells. ● Number is assigned to each antigen of the Rh system in order of
discovery
● D = Rh1 GENETIC WEAK D
● C = Rh2
● Complete expression of D antigen, but few in numbers
● E = Rh3
● c = Rh4
PARTIAL D (D MOSAIC)
● e = Rh5
WIENER FISHER-RACE ROSENFIELD ● Missing or altered epitope of D
r"Rz dcE/DCE Rh: 1, 2, 3, 4, -5 ● Produces anti-D (directed to missing epitope)
R2r DcE/dce Rh: 1, -2, 3, 4, 5
rr' dce/dCe Rh: -1, 2 -3, 4, 5 LW BLOOD GROUP SYSTEM__________
ryr" dCE/dcE Rh: -1, 2, 3, 4, -5 ● Demonstrated as antigen present on Rhesus monkey and on majority
R0ry Dce/dCE Rh: 1, 2, 3, 4, 5 of human RBC
● Originally thought to be the same antigen as Rh
INTERNATIONAL SOCIETY OF BLOOD TRANSFUSION (ISBT)
● 6-digit number ANTI – LW REACTION
● First three represent the system, the remaining for antigenic
specificity STRONG
FISHER-RACE WIENER ROSENFIELD ISBT
D Rh0 Rh1 004 001 ● With most Rh-positive RBC
C rh' Rh2 004 002
E rh” Rh3 004 003 WEAK
c hr' Rh4 004 004
r hr" Rh5 004 005 ● With Rh-negative RBC
NEGATIVE
RH TYPING__________________________
● Detects antigen only; NO REVERSE TYPING ● With Rh null cells
● Specimen: Red cells
● Reagent: anti-D typing sera; 22% BSA as control
● Weak reaction should be confirmed with Weak-D (IAT) Typing MAJOR BLOOD GROUP
● Weak D (Du) – Occurs when D is weakly expressed; more common
in black
LEWIS BLOOD GROUP SYSTEM (ISBT
WEAK D TYPING_____________________ 007)_________________________________
● Detects weak expression of D antigen on RBC surface
● Only blood group system not intrinsic to Red Blood Cell
● Follows principle of Indirect Antiglobulin Test (IAT)
● Made by tissue cells and secreted into body fluids -> adsorbed into
● Specimen: Negative test from Rh typing
RBCs
● Reagent: AHG (anti-human globulin)
● Two major antigens: Lea and Leb (not antithetical)
● Washing step: the most important step in AHG phase ● Lewis gene is located at chromosome 19p13.3 (Le) while 19q is Se
● Check Cells: “O positive” red cells coated with anti-D; added to a gene
negative anti – D test (Rh typing)
● Lewis antigen are poorly developed at birth
● Weak D positive test is reported as RH positive
● Leb antigen – receptor for Helicobacter pylori
● Development of Lewis Antigen: Le (a-b) – Le (a+b-) – Le (a+b+) –
CAUSES OF WEAK – D________________ Le (a-b+)
● Lewis Phenotypes
C TRANS TO RHD o Le (a+b) “NON-SECRETOR” - non-secretor of ABH but
still secrete Lea
● aka “Positioning Effect” Dce/Ce o Le (a-b+) “SECRETOR” – both Lea – soluble and Leb -
● D allele is in trans (opposite side) to the allele carrying C soluble antigens can be found in secretion and plasma
o Le (a-b-) “SECRETORS OR NON-SECRETORS” (lele) –
Del (D -) has nonfunctional Lewis transferase = no expression of
● No genes needed to synthesize Cc or Ee Lewis antigen on red cells
● Changes in Lewis Phenotypes: Pregnancy, Cancer, Viral/Parasitic,
RH NULL (--/--) Alcoholic Cirrhosis
● Lewis Antibodies are naturally occurring; IgM (cold reactive);
● No Rh antigens on its RBC considered insignificant in blood transfusion practices
● Mild compensated hemolytic anemia, stomatocytosis, ● Can be neutralized by Lewis substance present in secretions
hyperbilirubinemia, low haptoglobin, increased HbF, low Hb and
Hct RULES REGARDING LE ANTIGEN EXPRESSION
RH MOD ● 1. Lele individual will not produce any antigen: Le (a-b-)
● 2. A person with at least one Le gene and sese genes will be Le (a+b-)
● Partially suppressed Rh gene expression ● 3. A person who inherits at least one Le gene and at least one Se gene
● Similar to Rh null but milder will be leb positive: Le (a-b+)
F(ce)
● ‘f” antigen is expressed when both c and e in the cis position or
located in the same haplotype (e.g., Dce/DCE)
MNS BLOOD GROUP SYSTEM (ISBT I BLOOD GROUP SYSTEM (ISBT 027)___
002)_________________________________ I ANTIGEN
MN ANTIGENS ● “I” antigen stands for “individuality” and can be neutralized by
Human Milk
● Found in glycophorin A
● I is a public antigen; i antigen is found on cord blood cells; “I” =
● Exhibit dosage effect
adults; “i" = infants
● Well-developed at birth ● At birth: increased i antigen; I antigen is almost undetectable -> the
● M and N differ in amino acid residues at positions 1 and 5 quantity of i slowly decreases as I increases -> adult red cell rich in
● M = Serine and Glycine; N = Leucine and Glutamic Acid
I antigen; trace amount of i antigen
● Destroyed by enzyme treatment and ZZAP + DTT + papain or ficin
● “I” activity is increased in individuals with Bombay phenotype and
if ABH sugars are removed by enzymes
Ss ANTIGENS
● Found in glycophorin B I ANTIBODIES
● Well-developed at birth ● Autoanti – I – associated with Cold Agglutinin Syndrome;
● S and s differ in amino acid; S = methionine; s = threonine
Mycoplasma pneumonia infection (Primary Atypical Pneumonia)
● Less easily degraded by enzymes; variable effect from ficin
demonstrates high titer reactivity at 4oC and reacts over wide
treatment
thermal range (up to 30 – 32oC)
● Anti – I – saline reactive IgM auto agglutinin, detectable at 4oC
MNS ANTIBODIES o Benign anti – I: found in the serum of many normal healthy
● Anti – M = naturally occurring, saline agglutinin, cold reacting; individuals
reacts best at 6.5 pH ● Pathologic Anti -I: potent IgM agglutinins; reacting up to 30oC or
● Anti – M lectin = Iberis amara 32oC; causes agglutination and vascular occlusion or intravascular
● Anti – N – cold reactive – may be found in renal patients under hemolysis
dialysis on equipment sterilized with formalin ● Anti – i – associated with infectious mononucleosis (Epstein Barr
● Anti – N lectin – Vicia graminea, Bauhinia variegate, Bauhinia Virus)
purpura
● Anti – S and Anti – s – IgG in nature, reacts best at 37oC; can cause
severe HTR and HDFN
KELL BLOOD GROUP SYSTEM (ISBT
● U Phenotype – U stands for “universal”; U antigen is always present 006)_________________________________
when S or s is inherited; Anti – U is usually IgG and is formed by S- ● Antithetical antigens: K = Kell, k = Cellano; Kp a = Penny, Kpb =
s individual, usually of Black Origin (African) Rautenberg; Jsa = Sutter, Jsb = Matthews
● Originated from Mrs. Kellaher, from whom anti – K was first
P BLOOD GROUP SYSTEM (ISBT 003)__ identified which is most immunogenic second to D
● KEL gene found at chromosome 7
P1 ANTIGEN
KELL ANTIGENS
● Deteriorate rapidly on storage
● K is considered second most immunogenic next to D antigen
● P1 like antigens has been found in plasma, and droppings of pigeons
● Well developed at birth
and turtledoves, as well as in egg white
● K antigen = low frequency antigen
● P1 substance has been identified in hydatid cyst fluid, extracts of
● k antigen = high frequency antigen
Lumbricoides terstris, and Ascaris suum
● McLeod Phenotype – X – linked, occurs when Kx and km antigen
is not expressed and other Kell antigens are depressed
P ANTIBODIES
● Associated with Chronic Granulomatous Disease (CGD)
● 1. Anti – P1 – naturally occurring, IgM; weak, cold reactive saline ● NADH-oxidase is deficient in WBCs
agglutinin ● K-negative when transfused with K-positive will create anti-K as
o Neutralized by hydatid cyst fluid from Echinococcus high as 10% chance
granulosus infection, pigeon droppings or turtle dove egg white ● RBCs are acanthocytic; with compensated hemolytic anemia, and
● Anti – Tja (anti – PP1Pk) – discovered in serum of patient “Mrs. neurological and muscular abnormalities
Jay” w/ adenocarcinoma of stomach
o Associated with increased incidence of spontaneous abortions KELL ANTIBODIES
in early pregnancy
● Antibodies are warm reactive, IgG, reactive in AHG phase, involved
● Autoanti – P – associated with PCH (Paroxysmal Cold
in HDN and HTR
Hemoglobinuria)
● Anti – K – commonly encountered in blood bank; IgG; reactive in
o IgG antibody having a biphasic hemolysin Donath –
IAT phase
Landsteiner Test
● Made in response to antigen exposure through pregnancy and
o Antibody biphasic: binds RBC at cold temp (4oC) -> hemolyzes
transfusion
RBC at warm temp (37oC)
● Red cell destruction is usually extravascular
● Can cause severe HTR and severe HDFN
DISEASE ASSOCIATIONS
● P antigens – pyelonephritogenic E. coli, Streptococcus suis, Shigella
dysenteriae, Vibrio cholera, V. parahaemolyticus
● Receptor for parvoB19
● Anti – P1 – parasitic infections
JK null – abundant in Polynesians; also, been found in Filipinos,
DUFFY BLOOD GROUP SYSTEM (ISBT ●
Indonesians, Chinese, and Japanese
008)_________________________________ ● Organisms with JKb - like specificity include Enterococcus faecium,
Micrococcus, and Proteus mirabilis
DUFFY ANTIGENS
KIDD ANTIBODIES (ANTI JKa AND JKb)
● Fya and Fyb antigens – antithetical; expressed on cord blood;
destroyed by enzymes; receptor of Plasmodium vivax and ● Known to have a notorious reputation, associated with delayed
Plasmodium knowlesi HTR; causes infrequent and mild HDFN
● Fy (a-b-), Duffy null phenotype – is associated with African – ● Antibodies disappear rapidly both in vivo and in vitro
American population confers resistance to Plasmodium vivax ● Usually, IgG (warm reactive) and antibodies are labile on storage
infections ● Rh Null antibodies = anti – Jk-3
DUFFY ANTIBODIES LUTHERAN BLOOD GROUP SYSTEM

● Anti – Fya and Anti – Fyb are usually IgG (warm reacting), reacts
at AHG phase, can cause HTR, occasionally causes severe HDF
(ISBT 010)____________________________
● Antibody was first identified in a donor’s name, Luteran (label at the
● Anti – Fy3 – antibody made by Duffy Null phenotype
tube was misspelled)
● Can produce a mixed field agglutination
KIDD BLOOD GROUP SYSTEM (ISBT ● Anti – Lua reacts well with saline, can produce mixed – field
agglutination; thermal optimum is 12 – 23oC
009)_________________________________ ● Anti – Lub is an incomplete antibody; IgG – reacts better in AHG
● Located on RBC urea transporter phase; clinically significant
● Amorphic Gene = Lu (a-b-) while inhibitor gene = InLu, dominant
JKa AND JKb ANTIGENS type
● Well – developed at birth; enhanced in enzyme treatment; ● Antibody demonstrates a characteristic loose mixed-field
Demonstrate dosage effect agglutination pattern
● JK (a-b-) RBCs resist lysis in 2M urea
MINOR BLOOD GROUP

Blood Group System ISBT Notes
Diego Blood Group ISBT ● Location: anion exchange protein (AE-1) or also known as erythrocyte band 3
System 010 ● Linked to Mongolian Ancestry
● Mutation in AE-1 results to HS, Congenital Acanthocytosis, and Southeast Asian Ovalocytosis
Cartwright Blood ISBT ● Yta and Ytb; found in acetylocholinesterase
Group System 011
Xg Blood Group ISBT ● Location: X chromosome short arm
System 012 ● Destroyed by enzyme treatment
Scianna Blood Group ISBT ● Ag are expressed by the RBC adhesion protein ERMAP in humans
System 013
Dombrock Blood ISBT ● In PNH III – total absence of DO antigens
Group System 014
Colton Blood Group ISBT ● Location: Aquaporin – 1 (AQP)
System 015
Chido/Rodgers Blood ISBT ● Found in C4 Complement Components
Group System 017
Gerbich Blood Group ISBT ● Location: Glycophorin C and D; associated with RBC membrane band 4.1
System 020 ● Yus phenotype (Ge: -2, 3 4)
● Gerbich Phenotype (Ge: -2, -3, 4)
● Leach Phenotype (Ge: -2, -3, -4) presents with elliptpcytosis
Cromer Blood Group ISBT ● Location: Decay Accelerating Factor (CD55 or DAF)
System 021 ● PNH III – absence of Cromer antigen
Knops Blood Group ISBT ● Location: Complement Receptor 1 (CR1 or CD35)
System 022 ● Decreased expression in cases of SLE and chronic cold agglutinin disease
Indian Blood Group ISBT ● Location: CD44 glycoprotein
System 023
OK Blood Group ISBT ● Antigens carried on CD147; member of immunoglobulin superfamily
System 024
Raph Blood Group ISBT ● MER2 – only antigen; located on CD151, essential for assembly of basement membranes in the kidney and skin
System 025
John Milton Hagen ISBT ● Location: Semaphorin CDw108
(JMH) Blood Group 026 ● PNH III – absence of JMH
System ● Bennett – Goodspeed Antigens: Blood group associated with HLA (MHC Class III); Bga = HLA-b7; Bgb = HLA – B17;
Bgc = HLA – A28; Bg antibodies are destroyed by treating RBC antigens with chloroquine or EDTA glycine – HCL
● SID: Sda is associated with mixed-field refractile agglutination; neutralized by urine from an Sda positive individuals
and guinea pigs
SUMMARY__________________________ Venipuncture
site
No infectious skin disease or scars indicative of drug use
ANTIBODY CHARACTERISTICS
CLINICALLY SIGNIFICANT ABO, Rh, Kell, Kidd, Duffy, SsU, Lu b
USUALLY CLINICALLY I, Lewis, M, N, P, Lua ADJUSTMENT OF BLOOD VOLUME
INSIGNIFICANT
NATURALLY OCCURRING ABO, Lewis, P, MN, Lua AND BLOOD ANTICOAGULANT TO
WARM ANTIBODIES Rh, Kell, Duffy, Kidd DONORS WEIGHING <110 POUNDS____
COLD ANTIBODIES M, N, P
● 1. Convert kg to pounds by multiplying to 2.2
USUALLY ONLY REACT IN AHG Kell, Duffy, Kidd
PHASE ● 2. Determine allowable volume of blood to drawn (mL blood =
CAN REACT IN ANY PHASE Lewis weight in pounds x 450 mL / 110)
DETECTION ENHANCED BY Kidd, Lewis, I, Rh, P, ABO, (KLIRP – ABO) ● 3. Determine volume of anticoagulant needed (mL anticoagulant
ENZYME TREATMENT needed = mL blood / 100 x 14)
NOT DETECTED WITH ENZYME M, N, Duffy, Xga, Ch-Rg, JMH ● 4. Determine volume of anticoagulant to remove (mL anticoagulant
TREATMENT OF CELLS to be removed = 63 mL – mL anticoagulant needed)
(DESTROYED)
ENHANCED BY ACIDIFICATION M
SHOW DOSAGE Kidd, Duffy, Rh, MNS, Lutheran AUTOLOGOUS TRANSFUSION________
BIND COMPLEMENT Kidd, I, Lewis, Anti - Fya ● Criteria for autologous transfusion
CAUSE IN VITRO HEMOLYSIS ABO, Lewis, Kidd, Vel, some P o 1. Hemoglobin: > 11 g/Dl
LABILE IN VIVO AND IN VITRO Kidd o 2. Hematocrit: > 33%
COMMON CAUSE OF DELAYED Kidd o 3. NO BACTEREMIA
HTR ● For autologous transfusion, blood may be drawn from patient every
ASSOCIATED WITH Anti – P 3 days but NOT within 72 hours of scheduled surgery
PAROXYSMAL COLD
HEMOGLOBINURIA
ASSOCIATED WITH CAD AND Anti – I METHODS OF HEMOGLOBIN
PAP
ASSOCIATED WITH INFECTIOUS Anti – I DETERMINATION____________________
MONONUCLEOSIS ● 1. Copper Sulfate density
o Specific gravity of rgt: 1.053
o Acceptable drop of blood will sink in solution within 15
BLOOD DONOR AND SELECTION seconds
PROCESS o 30 mL copper sulfate = 25 tests
● 2. Spun microhematocrit
● The selection of potential blood donors and the subsequent ● 3. Spectrophotometric – most sensitive
collection and processing of those donor units are the first stages of
the blood banking process that eventually led to the transfusion of
lifesaving blood products to a patient TYPES OF BLOOD DONATION_________
● Donor screening encompasses the (1) medical history requirement ● 1. Allogeneic Donation – blood taken from individual of same
for the donor, (2) the (mini) physical examination, and (3) serologic species as the recipient
testing of the donor blood ● 2. Autologous Donation – the recipient is the donor, safer than
allogeneic
3. Apheresis – blood withdrawn from donor and separated into
THE DONATION PROCESS____________ ●
components, only the necessary component is harvested and
transfused
REGISTRATION
● Name, date and time of donation, address, telephone, gender, age or TYPES OF AUTOLOGOUS DONATION__
date of birth, consent to donate, additional information ● 1. Preoperative
o Occurs during the 5 to 6 weeks immediately preceding a
INTERVIEW AND APPEARANCE scheduled, elective surgical procedure unless the red blood cells
● The questionnaire is designed to be self-administered by the donor and plasma are scheduled to be frozen
but if preferred may be administered by a trained donor historian o Label must also clearly state “For Autologous Use Only”
● 2. Acute Normovolemic Hemodilution
o Results in the collection of whole blood with the concurrent
REQUIREMENTS FOR ALLOGENEIC infusion of crystalloid or colloid solutions, thus maintaining a
DONATION__________________________ normal blood volume but decreasing the patient’s hematocrit
● Treat all specimens such as blood, body fluids and unfixed tissues to o Ratio of replacement is 3:1 for crystalloids and 1:1 for colloids
be potentially infectious ● 3. Intraoperative Collection
PARAMETER PHILIPPINE STANDARD AABB STANDARD o Involves collecting shed blood from the surgical site;
General Appears to be healthy processing the blood through an instrument that washes it with
appearance saline to remove tissue debris, free hemoglobin, and plasma that
Age 16-65 years old; 16-17 y/o with At least 17 years old may contain activated coagulation factors; concentrating the
guardian consent; >65 y/o (minors with consent, residual red cells (to a hematocrit of 50% to 60%); and then
blood bank physician will elderly with blood bank reinfusing those cells immediately
decide physician’s consent) ● 4. Postoperative Blood Salvage
Weight >50 kg or >110 lbs o Collected from a drainage tube placed at the surgical site
Temperature Does not exceed 37.5oC or 99.5oF
o It is reinfused, with or without processing, via a microaggregate
Pulse 50 – 100 beats/min (60 – 100 beats/min in some reference)
filter to screen out any debris
Blood Pressure Systolic: 90 – 160 mmHg Systolic: <180 mmHg
Diastolic: 60 – 100 mmHg Diastolic: <100 mmHg o This blood is characterized as being dilute, partially hemolyzed,
Hemoglobin >12.5 g/dL and defibrinated
INDEFINITE DEFERRAL
DEFERRAL GUIDELINES FOR
ALLOGENEIC DONATION____________ ● Prospective donor is unable to donate blood for someone else for an
unspecified period of time due to current regulatory requirements
DURATION CAUSE
2 DAYS Aspirin (if donor is sole source of platelets) PERMANENT DEFERRAL
2 WEEKS ● Vaccination for measles, mumps, polio, typhoid,
yellow fever ● Prospective donor will NEVER be eligible to donate blood for
● After febrile illness (fever) someone else; may donate autologous blood (Confirmed cases of
6 WEEKS ● Pregnancy after delivery infection, severe clinical conditions, and very high risk of infection)
● Abortion/miscarriage without dilatation and curettage
(1 year if with dilatation or curettage)
1 MONTH ● Vaccination for Rubella and chicken pox
BLOOD COLLECTION________________
● Proscar, Propecia, and Accutane
2 MONTHS Whole blood donation COLLECTION OF WHOLE BLOOD
6 MONTHS Avodart Skin preparation Aseptic method (iodine and quats; chlorhexidine
1 YEAR ● Syphilis gluconate; 70% isopropyl alcohol) – scrub site at least
● Gonorrhea 4 cm in all direction for 30 seconds
● Mucous membrane exposure to blood (e.g., tattoo)
Torniquet/ 40 – 60 mmHg
● Skin penetration with sharp contaminated with blood
or body fluids
Sphygmomanometer
● Household or sexual contact with HIV at a high risk Needle Blood donation = Gauge 16; blood transfusion =
● Incarceration in correctional facility for > 72 Gauge 18; Angle: 20 degrees
consecutive hour Volume of blood 450 mL +/-10% or 500 mL +/-10%
● Travel to Iraq or area endemic for malaria (e.g., routinely collected
Palawan) Maximum volume 10.5 mL of blood per kilogram of donor’s weight (450
● Recipient of blood, blood components, plasma – mL blood + 30 mL in diversion pouch for testing;
derived clotting factor concentrates, or transplant maximum total of blood collected = 525 mL)
(after bite from an animal) Low volume 300 – 404 in 450 mL bag or 333 – 449 mL in 500 mL
● Rape victims collections bag (labeled as “low-volume”); RBCs may be
● After therapeutic rabies virus transfused but other components SHOULD NOT BE
● After hepatitis B immune globulin administration PREPARED
3 YEARS ● Malaria, or from an area endemic for malaria
Volumes of 63 mL for 450 L blood; 70 mL for 500 mL blood;
● Soriatane
anticoagulant Anticoagulant to blood ratio = 1:8
PERMANENT ● Parenteral drug use
● Family history of Creutzfeldt-Jakob disease
Time of collection Usually <10 minutes; if >15 minutes, unit may not be
● Treatment with pituitary growth hormone of HUMAN suitable for preparation of PLATELETS, FRESH
ORIGIN FROZEN PLASMA, AND CRYOPRECIPITATED
● Viral hepatitis after 11 th birthday ANTIHEMOPHILIC FACTOR (AHF)
● Confirmed positive HBsAg Sample for testing From diversion pouch or by 2nd phlebotomy
● Repeatedly reactive anti-HBc on greater than 1 Storage temperature For platelets = 20 – 24oC; Other components = 1 – 6oC
occasion of unit between
● Repeatedly reactive HTLV on greater than 1 occasion collection and
● Present or past clinical or laboratory evidence of processing
infection with HIV, HCV, HTLV
● History of Babesiosis or Chagas’ disease SUBMISSION OF WHOLE BLOOD TO BLOOD BANK / CENTER
● Person who has engaged in sex for money or drugs
anytime since 1977 ● Collected units stored at 1 – 6oC, submitted within: 24 hours
● Men who have had sex with another man anytime ● Collected units stored at 20 – 24oC, submitted within: 6 – 8 hours
since 1977 ● Whole blood submitted within 6 -8 hours after collection can be used
● Hemophiliacs for platelet preparation
● Person with chronic cardiopulmonary, liver, and renal
disease LABELLING OF BLOOD BAGS
● Chemotherapeutic agents administered for
malignancy FDA 1985 RA 1517
● Hematologic malignancies BLOOD TYPE COLOR LABEL BLOOD TYPE COLOR LABEL
● Serious abnormal bleeding tendencies A Yellow A Blue
● Those who have taken TEGISON (Etretinate) B Pink B Yellow
NO Toxoid vaccine (killed/synthetic viral, bacterial, rickettsial AB White AB Pink
DEFERRAL without symptoms) O Blue O White
Treated and inactive TB patient “YES Po Whole Blood” “BY PassWord”
Recombinant growth hormone
DONOR BLOOD UNIT PROCESSING
TYPES OF DEFERRAL ● All donor units are processed before being released for compatibility
testing and transfusion
TEMPORARY DEFERRAL ● Tests done to donor blood:
o 1. ABO grouping
● Prospective donor is unable to donate blood for a limited (specific) o 2. Rh typing
period of time o 3. Antibody screen (required only on those with previous
pregnancy and/or transfusion)
o 4. Screening test for transfusion – transmissible infections
BLOOD COMPONENT PREPARATION SEPARATION OF COMPONENT_______
● Within 6 – 8 hours
OPEN SYSTEM AND CLOSE SYSTEM__

SYSTEM EXPLANATION EFFECT ON
EXPIRATION DATE OF
COMPONENT
OPEN ● Seal on unit is broken ● Component stored at
SYSTEM to attach external 1 – 6oC must be used
transfer bag within 24 hours after
● Exposure to air poses system opened
threat of bacterial ● Components stored
contamination at 20 – 24oC within 4
hours
CLOSE ● Sterility maintained ● No change (retain
SYSTEM through use of attached original expiration
satellite bags or sterile date)
connecting device that
welds tubing from 1
bag to another
ERRATUM: CRYOPRICIPATATE change to CRYOPRECIPITATE ● No exposure to air
COMPONENTS______________________________________________________________
Blood Component Preparation and Indication Storage and Transport and Others
Shelf Life
1. Whole Blood Indication: Provide blood volume expansion and RBC Storage: 1 – 6 degrees Hct: 3% - 5% increase per unit
mass in acute blood loss; for active bleeding patients who Celsius Hgb: 1 – 1.5 g/dL increase per unit
have lost at least 25% of their blood volume or patients Transport: 1 – 10 degrees When whole blood is not available, reconstitute
requiring exchange transfusion Celsius whole blood by mixing RBCs with thawed AB
Shelf Life: ACD and CPD type plasma from different donor
= 21 days
CPDA-1 = 35 days
2. Packed RBCs Preparation: 80% of plasma removed from whole blood; Storage: 1 – 6 degrees Hct: 3% - 5% increase per unit
hematocrit: 65 – 80% (not exceed 80%) Celsius Hgb: 1 – 1.5 g/dL increase per unit
Indication: Indication of RBC mass symptomatic, Transport: 1 – 10 degrees
normovolemic patients; for oncology patients Celsius
undergoing chemotherapy or radiation therapy, trauma Shelf Life: Open system:
patients, dialysis patients, premature infants with sickle 24 hours; Close system:
cell anemia ACD and CPD = 21 days;
CPDA-1 = 35 days
3. Leukocyte – Preparation: Filtration (within 48 hours from time of Storage: 1 – 6 degrees
Reduced RBCs collection) or apheresis; saline washing; must retain 85% Celsius
of original RBCs; 1 unit contains 5 x 106 WBCs Transport: 1 – 10 degrees
Indication: Increase RBC mass in patients with severe Celsius
and/or recurrent febrile transfusion reactions due to Shelf Life: Open system:
leukocyte antibodies 24 hours; Close system:
● Increased RBC mass in patients at risk for HLA ACD and CPD = 21 days;
alloimmunization or susceptible to cytomegalovirus CPDA-1 = 35 days
(CMV)
4. Washed RBCs Indication: Increased RBC mass of symptomatic anemia Storage: 1 – 6 degrees NOT a substitute for leukoreduced RBCs; about
patients with transfusion history of allergic, urticarial Celsius 10 – 20% of RBCs lost in process using saline
reaction, anaphylactic reaction, febrile nonhemolytic Shelf Life: 24 hours
reaction
● Used in infant or intrauterine transfusion
● 75% hct; <5 x 108 WBCs
5. Irradiated Preparation: Recommended minimum dose of gamma Storage: 1 – 6 degrees For prevention of graft versus host disease; kills
RBCs irradiation is 25 – 35 Gy Celsius donor T cells
● Minimum dose: center unit: 25 Gy; other parts of Shelf Life: Original
the unit; 15 Gy outdate or 28 days from
Indication: Immunodeficiency, malignancy, bone irradiation, whichever
marrow transplant, transfusion with blood from blood of comes first
relative, intrauterine and neonatal transfusion
6. Frozen RBCs Preparation: Frozen in glycerol within 6 days of Storage: Frozen: - 65 Osmolality to monitor glycerol removal; virtually
collection degrees Celsius or -20 all plasma, anticoagulant, WBCs, and platelets
Indication: Storage of rare blood and autologous unit degrees Celsius removed; safe for IgA-deficient patient
Shelf Life: 10 years; after
deglycerolization: 24
hours (unless closed
system)
7. Platelet Preparation: Centrifugation of whole blood at room Storage: 20 – 24 degrees 1 unit contains > 5.5 x 1010 platelets
Concentrate / temperature, within 8 hours of collection -> 1st light spin Celsius ● 40 – 70 mL plasma; pH = > 6.2
Random Donor yields platelet – rich plasma -> 2nd heavy spin separates Shelf Life: 5 days from ● One unit should increase platelets by 5,000 –
Platelet platelets from plasma collection with constant 10,000/uL in 75 kg recipient
Indication: Main indicator for use: if platelet count is less agitation; after pooling: 4 CORRECTED COUNT INCREMENT FOR
than 20,000/uL; Pre-operation platelet count is less than hours (open system) PLATELETS
50,000/uL a. Good increments: > 10,000 u/L
For bleeding due to thrombocytopenia or b. Refractoriness: <50,000/uL
thrombocytopathy; for patients with chemotherapy, post- PLATELET RECOVERY
bone marrow transplant patients, post-operative bleeding ● 60% at 1 hour
● NOT indicated in patients with idiopathic ● 40% at 25 hours
thrombocytopenia (ITP)
8. Plateletpheresis Preparation: Apheresis Storage: 20 – 24 degrees 1 unit contains > 3.0 x 1011 patients; equivalent to
Unit or Single Indication: For thrombocytopenic patients Celsius 4 – 6 hours
Donor Platelet alloimmunized to HLA or platelet antigen (donor should Shelf Life: 5 days with ● 200 – 400 mL plasma; pH = > 6.2
(SDP) be HLA matched) constant agitation ● 1 unit should increase platelets by 30,000 –
● Limit the donor exposure in thrombocytopenic 60,000/uL in 75-kilogram recipient
patients who acquired long term platelet ● Exposes recipient to fewer donors
transfusions\
9. Leukocyte – Preparation: WBCs removed by filtration or during Storage: 20 – 24 degrees -
Reduced apheresis processing Celsius
Platelets Indication: Same with Single Donor Platelet (SDP) or Shelf Life: Open system: 4
Plateletpheresis unit hours; Apheresis: 5 days
with agitation
10. Granulocyte Preparation: Apheresis; uses hydroxyethyl starch (HES) Storage: 20 – 24 degrees 1 unit contains > 1.0 x 1010 granulocyte, platelets,
as sediment agent Celsius WITHOUT and 20 – 50 mL of RBCs
● Administer corticosteroid to donor 12 – 24 hours agitation PLASMA DERIVATIVES
before donation Shelf Life: 24 hours ● Main indication for use: (1) PT is > 16
Indication: Main indication for use: For platelet with seconds (INR 1.5), (2) APTT is >60 seconds,
granulocyte count of <350/uL or <500/Ul (3) Fibrinogen is <100 mg/dL
Patients with granulocyte dysfunction or myeloid
hypoplasia who are unresponsive to antibiotics; severe
neutropenia with infection non-responsive to antibiotic
therapy
● Limited to septic infants
11. Fresh Frozen Preparation: Plasma separated from whole blood within Storage: Frozen: -18 ● Contains all coagulation factors; check for
Plasma 8 hours of collection degrees Celsius (1 year); evidence of thawing and refreezing; thawed
● FFP -> thaw in waterbath at 30 – 37 degrees Celsius after thawing: 1 – 6 at 30 – 37 degrees Celsius for 30 – 45 minutes
for 30 to 45 minutes -> thawed plasma (transfuse degrees Celsius for 24 or by FDA-approved microwave
immediately; store at 1 to 6 degrees Celsius up to 6 hours
hours or store at 4 degrees Celsius u to 24 hours if Shelf Life: Frozen: -18
factor VIII is not needed) degrees Celsius (1 year);
Indication: Bleeding patients who require factors II, V, after thawing: 1 – 6
VII, IX, X degrees Celsius for 24
● Replace isolated factor deficiencies when specific hours
component is not available
● Reverse effects of Warfarin
● Thrombotic thrombocytopenic purpura (TTP) and
Hemolytic Uremic Syndrome (HUS)
● Patients with liver disease to prevent or correct
bleeding
● Antithrombin II deficiencies; disseminated
intravascular coagulation when fibrinogen is < 100
mg/dL
12. Cryoprecipitate Preparation: Prepared by thawing FFP at 1 – 6 degrees Storage: Frozen: -18 Used for hemophilia A and von Willebrand’s
Celsius, removing plasma, and refreezing within 1 hour degrees Celsius; after disease ONLY IF factor VIII concentrate or
FFP -> thaw at 4 degrees Celsius -> centrifuge: Heavy thawing: room recombinant factor preparations not available
spin -> white mass of precipitate temperature
Contains: Shelf Life: After thawing:
1. Fibrinogen – 150 – 22 mg 6 hours (single units); 6
2. AHF VIII:c – 80 – 120 IU hours (pooled units, closed
3. vWF – 40 – 70% system), 4 hours (pooled
4. Factor XIII – 20 – 30% units, open system)
5. Fibronectin
Before infusion: Frozen cryoprecipitate -> thaw at 30 –
37 degrees Celsius -> thawed cryoprecipitate
For thawed cryoprecipitate: store at room temperature
and transfuse immediately; or transfuse within 6 hours
For pooled cryoprecipitate (after thawing): transfused
within 4 hours (open system)
Indication: For treatment of fibrinogen deficiency,
hemophilia A, von Willebrand’s disease and Factor XIII
deficiency, and as a fibrin sealant
● Factor VIII deficiency: routinely treated with Factor
VIII concentrate
● NOT indicated for thrombotic thrombocytopenic
purpura (TTP)
13. Plasma Preparation: Plasma may be separated from whole blood Shelf Life: 5 years when PLASMA EXPANDERS – products that are
Derivatives at any time during the unit’s shelf life up to 5 days after stored between 1 and 6 transfused in patients suffering from hypovolemia
the expiration degrees Celsius or indicated among burn and shock patients
Albumin, Plasma Protein Fractions (PPF), Immune 1. Plasma-derived volume expander
Serum, freeze dried Factor VIII, Freeze dried Factor IX • Normal Serum Albumin (NSA): 96%
Treated with different methods such as pasteurization, albumin + 4% globulin
nanofiltration, and solvent detergent to ensure sterility • Plasma Protein Fraction (PPF)l 83%
Lyophilized or freeze dried albumin + 17% globulin
2. Synthetic volume expander
• Crystalloids
a. Ringer’s Lactate (Na, Cl, K, Ca,
lactate ions)
b. Normal Saline Solution (0.85 –
0.90% NaCl)
• Colloids
a. Dextran (6 – 10%)
b. Hydroxyethyl starch (HES)
SUMMARY__________________________________________________________________
COMPONENT SHELF-LIFE STORAGE QUALITY VOLUME INDICATIONS FOR CONTENT DOSAGE TRANSFUSION
TEMP CONTROL USE CRITERIA
Whole Blood CPD 21 d 1 – 6oC 450 – 500 Volume expansion, RBC, Increased ABO, Rh
CPDA-1 35 d mL increased oxygen Plasma, hgb 1 g/dL
CP2D 21 d Platelets, Increased hct
WBCs 3%
Whole Blood Original 1 – 6oC 25 Gy to 450 – 500 Prevent CVHD volume RBC. Increased ABO, Rh
irradiated expiration or 28 d center of mL expansion, increased Plasma, hgb 1 g/dL
post-irradiation canister oxygen Platelets Increased hct
3%
RBCs CPD 21 d 1 – 6oC No 250 – 300 Increased oxygen RBC Increased ABO, Rh
CPDA-1 35 d additive: mL hgb 1 g/dL
CP2D 21 d hct < 80% Increased hct
ACD 21 d Additive: 3%
AS 42 d N/A
RBCs aliquots Closed system: 1 – 6oC Varies Increased oxygen RBC 10 mL/kg ABO, Rh
same Increased
Open system: 24 hgb 2 g/dL
hr
RBC irradiated Original outdate 1 – 6oC 25 Gy to 250 – 300 Prevent CVHD, increased RBC Increased ABO, Rh
or 28 d post- center of mL oxygen hgb 1 g/dL
irradiation canister Increased hct
3%
RBC Closed system: 1 – 6oC <5 x 106 250 – 300 Febrile run, increased RBC, Few Increased ABO, Rh
leukoreduced same WBCs mL oxygen platelets, hgb 1 g/dL
Open system: 24 >85% RBC plasma Increased hct
hr recovery 3%
Washed RBCs 24 hr 1 – 6oC HCT 70 – 180 mL IgA – negative persons RBC, Increased ABO, Rh
80% WBC < 5 = hgb 1 g/dL
109 Increased hct
3%
Frozen RBCs 10 years < –65oC Rare phenotypes RBC,
Glycerol
RBC 24 hr 1 – 6oC 80% of 180 mL Rare phenotypes, RBC, Increased ABO, Rh
deglycerolized RBC increased oxygen Saline, hgb 1 g/dL
recovery Dextrose < Increased hct
<1% 1%, WBC, 3%
glycerol Platelets
Platelets, 5–7d 20 – 24oC >5.5 x 1010 50 – 70 Thrombocytopenia Platelets Increased 5k
whole-blood plts mL – 10k/uL
derived (RD) pH > 6.2
Platelets, 5–7d 20 – 24oC > 3 x 1011 200 - 400 Platelet retractoriness Platelets Increased HLA compatible
apheresis (SD) plts mL 30k – 60k/uL
pH > 6.2
Platelets, 5d 20 – 24oC 25 Gy to Same Prevent CVHD Platelets Same Same
irradiated center of
canister
Platelets, 4 hr 20 – 24oC pH > 6.2 Varies Thrombocytopenia, DIC Platelets Varies
pooled bleeding
Platelets, 5d 20 – 24oC RD: <8.3 x pH > 6.2 Febrile runs Platelets RD: Increase SD: HLA
leukoreduced 105 WBCs 5 – 10%
SD: <5 X SD:
106 WBCs Increased 30
– 60%
FFP 1 yr –18oC 8 hr CPD, 200 – 250 Coagulation deficiency, 1U/mL Increased ABO
7 yr –65oC CPDA-1, mL Liver disease, DIC, clotting Factor 20 –
CP2D 6 hr Massive trx factors 30%
ACD 10 – 20
mL/kg
AFFP 1 yr < –18oC
PF24 1 yr –18oC 24 hr WB 150 – 250 Same Decreased Same Same
7 yr –65oC mL labile
factors
SDP 1 – 6oC liquid 5 days after Frozen: 6 150 – 250 Stable Same Same
WB hr mL clotting
expires factors
Frozen: 5
yr
LP 5 days after WB 1 – 6oC 200 – 380 Not well
expires liquid mL characterized
Cryoprecipitate Frozen: 1 yr –18oC FVIII:C: 80 10 – 25 Hemophilia A, VWD: FVIII:C Increased ABO
Thawed: 6 hr 20 – 24oC IU mL FXII deficiency, Fibrin (80 – fibrinogen
Pooled: 4 hr sealant, 120U) 5 – 10 mg/dL
Hypofibrinogenemia VWF (40 –
70%)
FXII (20 –
30%)
Fibrinogen
(150
mg/dL)
FVIII Check vial 1 – 6oC 10 - 30 mL Hemophilia A FVIII 1U FVIII/kg Reconstitute
concentrates Trace other body wt. before infusion
clotting Increase 2%
factors
FIX Check vial 1 – 6oC 20 – 30 Hemophilia B FIX 1U FIX/kg Reconstitute
concentrates mL Trace other body wt. before infusion
clotting Increase
factors 1.5%
Cryo-reduced 1 yr < –18oC 200 – 380
plasma mL
Granulocytes 24 hr 20 – 24oC > 1 x 1010 200 – 600 Neutropenia < 500 WBC, 1 – 2 x ABO, Rh, HLA
mL PMNs/uL RBC, 1010/infusion
Platelets, four daily
Plasma doses
Granulocytes, 24 hr 20 – 24oC > 1 x 1010 200 – 600 Prevent CVHD, Same Same Same
irradiated mL neutropenia
ISG 3 yr, IM Varies Prophylaxis, Gamma IM or IV
1 yr, IV Immunodeficiency, globulins
Hypogammaglobulinemia IgG, IgM,
IgA
NSA 5 yr, 25% 2 – 10oC 50 mL Plasma volume expansion
250 mL
PPF, 5% 5 yr 2 – 10oC 250 mL Plasma volume expansion
Dextran 6% (dex 40) Volume expansion burns
10% (dex 70)
HE5 6% Volume expansion burns
o
RhIg 2 yr 1–6 C 1 mL Rh HDN Anti – D 300 ug Mother Rh-
IgG 120 ug negative; baby
50 ug Rh-positive, Rh-
unknown
Intermittent – flow centrifugation – one venipuncture site, in

TRANSFUSION_______________________ o
which blood is withdrawn and reinfuse to the same needle
● The administration of blood or its component intravenously is done o Continuous – flow centrifugation – two venipuncture sites are
within 4 hours (2 – 4 hours) necessary
● At a rate of 60 drops per minute, 240 mL of blood can be transfused
● b. Filtration – removal of only plasma through a membrane;
within 1 hour
therapeutic process
● Time limit for infusion of blood units: ● c. Adsorption – removal of only a selected constituent of plasma
Blood Start of infusion Complete infusion with reinfusion of plasma after constituent is removed
Whole blood/PRBC Within 30 minutes of Within 4 hours
removing from ref FREQUENCY OF DONATION
Platelet concentrate Immediately Within 20 minutes
Fresh frozen plasma ASAP after thawing Within 20 minutes ● 1. Red cell pheresis – every 16 weeks
● 2. Plateletpheresis – 48 hours interval, not more than 24 times a year
HEMAPHERESIS / APHERESIS ● 3. Plasmapheresis – every 4 weeks (total protein, IgG, and IgM
monitored)
● 4. Leukopheresis – not more than twice a week, not more than 24
METHODS times a year
● a. Centrifugation – withdrawal of whole blood, removing selected

fraction and reinfusion of the remaining components into the door
TRANSFUSION REACTION
● 1. Immediate – immunologic / non-immunologic
● 2. Delayed - immunologic / non-immunologic
IMMEDIATE - IMMUNOLOGIC
Signs and Symptoms and Management
1. Acute Hemolytic ● Reaction due to clinically significant antibody reacting Signs and Symptoms
Transfusion Reaction at 37 degrees Celsius Fever, chills, flushing, nausea, dyspnea, chest pain, flank pain,
(AHTR) ● Will result to red cell destruction (intra/extravascular hypotension, shock, hemoglobinemia, hemoglobinuria, DIC, renal
hemolysis) failure
Management
Adequate renal perfusion, induce – diuresis, treat shock and manage
disseminated intravascular coagulation (DIC)
2. Febrile Non- ● Reaction is due to patient’s anti-leukocyte antibody Signs and Symptoms
Hemolytic ● Increase in temperature of 1 degree Celsius or more that Chills, fever
is associated with transfusion Management
Administer antipyretics
3. Allergic ● Reaction is due patient’s anti-plasma protein antibody Signs and Symptoms
Hives
Management
Administer antihistamine while blood flow is slowed or stopped
Prevention: Washed RBC
4. Anaphylactic ● Reaction is due to patient’s anti-IgA Signs and Symptoms
● Occur only after the infusion of only few mL of blood Flushing of the skin, abrupt hypertension followed by hypotension,
● NO FEVER substernal pain, dyspnea, nausea, abdominal cramps, emesis, diarrhea
Management
Give immediate treatment with epinephrine, IV corticosteroids and
oxygen therapy may be indicated
Prevention: Washed RBC
5. Transfusion Related ● Reaction is due to leukoagglutinins Signs and Symptoms
Acute Lung Injury Chills, fever, nonproductive cough, dyspnea, cyanosis, bilateral
(TRALI) / Non- pulmonary edema, sever hypoxemia, hypotension
Cardiogenic Management
Pulmonary Edema Give respiratory support, steroids and diuretics
(NCPE) Prevention: Leukocyte-reduced RBC
IMMEDIATE NON-IMMUNOLOGIC
1. Transfusion – ● Principal causes are transient bacteremia in Signs and Symptoms
Associated Sepsis asymptomatic donors and also contamination of Fever, chills, hypotension, tachycardia, shock, hemoglobinemia,
collection equipment during the manufacturing process hemoglobinuria, renal failure, DIC
● Common bacteria isolated (Psychrophilic bacteria) Management
● Yersinia enterocolitica: MOST COMMON Give IV antibiotics, fluids, and vasopressors to maintain blood
● Pseudomonas spp. pressure, appropriate therapy for DIC (if present)
2. Transfusion – ● Associated with rapid infusion of large volumes of Signs and Symptoms
Associated blood products Anxiety, restlessness, coughing, tachycardia, dyspnea, cyanosis,
Respiratory Overload ● At risk: children, elderly, cardiac disorder px severe headache, signs of congestive heart failure, peripheral edema
(TACO) Management
Administer diuretics, place patients in upright position (O2 by mask,
IV morphine, phlebotomy of 200 – 400 mL of blood if necessary)
3. Physical or Chemical ● Infusion of blood in a small-bore needle, addition of Signs and Symptoms
Hemolysis hypo/hypertonic solution, and thermal trauma Asymptomatic hemoglobinuria (hemoglobinemia, DIC, and renal
failure as RARE)
Management
Generally, none needed
DELAYED – IMMUNOGENIC
1. Delayed Hemolytic ● One cause is Kidd antibody Signs and Symptoms
Transfusion Reaction Fever, decreased hemoglobin, jaundice
(DHTR) Management
Treatment is rarely necessary, give antigen – negative blood for
subsequent transfusion
2. Transfusion – ● Occurs when certain susceptible recipient with Signs and Symptoms
Associated VS Host compromised immune systems is transfused with blood Acute: fever, diffuse skin rash, diarrhea infection, abnormal liver
Disease or blood component containing immunocompetent function, pancytopenia, usually fatal
lymphocytes Chronic: fever, scleroderma-like disease, Sicca syndrome, interstitial
pneumonitis, malabsorption
Management
No adequate therapy
Prevention: Irradiation using 137Ce or 60Co
DELAYED NON-IMMUNOGENIC
1. Hemosiderosis ● Excess iron accumulates in mitochondria of cell in Signs and Symptoms
organs such as the liver, heart, and endocrine gland Muscle weakness, weight loss, mild jaundice, fatigue, cardiac
● At risk px: Thalassemia major, Sickle cell anemia, arrythmias, mild diabetes, growth retardation in children
hemoglobinopathies Management
Administering deferoxamine
2. Disease Transmission ● Hepatitis B, C, D Signs and Symptoms
● CMV, EBV, HIV Fever, fatigue, lymphadenopathy, adenopathy, malaise, arthralgia,
● T. pallidum, Plasmodium spp. icterus, hemolysis
Management
Notify facility for drawing blood, quarantine components in storage
prepared from same unit
cell adherence or
TRANSFUSION – TRANSMITTED particle agglutination
DISEASES T. pallidum antigen –
specific
● 10 tests include:
immunofluorescence or
o 1. HbsAg agglutination assays
o 2. HBc antibody
Trypanosoma ChLIA or EIA Radioimmunoprecipitation
o 3. HCV antibody; HCV NAT testing
cruzi assay (RIPA)
o 4. HIV ½ antibody
o 5. HIV-1 p24 antigen; HIV-NAT testing
o 6. HTLV-I/II antibody BLOOD BANKING TECHNIQUES
o 7. Syphilis
o 8. Cytomegalovirus
o 9. Trypanosoma cruzi antibody/Chagas Disease ANTIBODY SCREENING______________
o 10. West Nile Virus NAT ● Detects alloantibodies/unexpected antibodies
● Involves reaction between patient serum or plasma with 2 – 3
Disease Method Confirmatory reagents
HIV 1/2 Chemiluminescent Immunofluorescence assay ● Source of antigens: reagent antibody screening cells
(ChLIA) or Enzyme (IFA) or Western blot ● Source of antibodies: patient’s serum
Hepatitis B Immunoassay (EIA) Neutralization
Hepatitis C Recombinant Immunoblot
Assay (RIBA)
Syphilis Microhemagglutination
or EIA: solid-phase red
ANTIBODY IDENTIFICATION / PANEL TESTING____________________________
● More definitive test to determine antibodies are present in the serum if antibody screen is positive
● Source of antigen: Reagent antibody panel cells (10 – 16 cells)
● Source of antibodies: patient’s serum
CROSSMATCHING___________________ TYPES OF CROSSMATCH

● Determines compatibility of donor’s RBCs with recipient blood ● a. Immediate spin – recipient serum and donor RBCs tested in
immediate spin only; detects ABO, antibody screen
MAJOR CROSSMATCHING ● b. Antiglobulin – recipient serum and donor RBCs tested through
AHG phase
● Consist of mixing the RECIPIENT’S SERUM + DONOR’S RED
● c. Computer – Computer check of donor and recipient ABO and Rh
CELL
type
● Source of antigen: Donor cell ● d. Abbreviated – type and screen with immediate spin crossmatch
● Source of antibodies: Recipient’s serum
● e. Broad spectrum – most preferred; has 3 phases: (1) immediate
spin, (2) 37 degrees Celsius phase, and (3) Antihuman globulin
MINOR CROSSMATCHING
phase
● Consist of mixing the DONOR’S SERUM + RECIPIENT’S RED
●
CELL
Source of antigen: Recipient’s red cells
SEROLOGIC TECHNIQUES___________
● a. Enzyme Technique – use of enzyme treated RBCs to enhance or
● Source of antibodies: Donor’s serum
remove reactivity of some antibody specificities
● b. Elution – technique used to dissociate IgG antibodies from
sensitized RBCs
c. Adsorption – process of removing antibody from serum by
●
combining a serum sample with appropriate RBCs
RH HDFN____________________________
● Rh-positive firstborn infant of an Rh-negative mother usually
● d. Neutralization – uses soluble antigen to inhibit reactivity of
unaffected because the mother has not yet been immunized
certain antibodies in hemagglutination assays
TECHNOLOGIES IN BLOOD BANKING

TUBE TESTING
● Gold standard; glass test tubes used as reaction container; observed
for agglutination and/or hemolysis
GEL TECHNOLOGY
● Uses plastic microtube containing dextran acrylamide gel as reaction
container
● Tests Applicable: ABO blood grouping, Rh typing, DAT, antibody
screening, antibody identification, and compatibility testing
● Observe for agglutinated RBCs suspended in gel (positive reaction;
graded)
● Advantages: STANDARDIZATION, stability, decrease sample
volume needed for testing, enhanced sensitivity and specificity
● Disadvantages: need special incubator and centrifuge; specific
pipette for 25 microliter of serum or plasma and 50 microliter of
0.8% RCS
SOLID PHASE TECHNOLOGY

● Test reactant is bound to solid support (microtiter wells) before test
is started
● Positive: microplate with RBC membranes bound to surface of wells
● Negative: no adherence of RBCs in bottom of well
AFFINITY COLUMN TECHNOLOGY

● Principle of affinity adherence of IgG sensitized RBCs to an
immunologically active matrix
DURING GESTATION AND DELIVERY
HEMOLYTIC DISEASE OF THE ● Fetal RBCs enter maternal circulation -> When D antigen is
inherited from the father, fetal cells immunize the mother and
FETUS AND NEWBORN stimulate the production of anti-D (all subsequent offspring who
● Hemolytic Disease of the Newborn inherit D antigen will be affected) -> Maternal anti-D cross the
o Caused by the destruction of the RBCs of the fetus by antibody placenta and bind to the fetal Rh-positive cells -> Sensitized RBCs
produced by the mother (IgG) are destroyed (hemolyzed) by fetal monocyte – macrophage system,
o Only antibodies of the immunoglobulin G (IgG) class are resulting in ANEMIA
actively transported across the placenta
o Maternal antibody developed during: (1) Previous pregnancy, FACTORS AFFECTING IMMUNIZATION AND SENSITIVITY OF
(2) Previous transfusion, (3) Second or third trimester HDFN
o 95% of HDFN cases caused by maternal antibodies against Rh
(D) antigen ● 1. Antigenic Exposure
o Fetomaternal hemorrhage -> increase in maternal antibody
titers -> increase severity of HDFN
DIAGNOSIS AND TREATMENT OF HDFN o As little as 1 mL of fetal RBCs can immunize the mother
● Rh-immune globulin (RhIg) ● 2. Host Factors
● Ultrasonography o In Rh-negative individuals who are transfused with 200 mL of
● Doppler assessment of middle cerebral artery peak systolic velocity Rh-positive RBCs, approximately 85% respond and form anti-
● Cordocentesis D
● Allele-specific gene amplification studies in fetal cells in amniotic o Nearly all of the nonresponders will fail to produce anti-D even
fluid with repeated exposure to Rh-positive blood
● Fetal DNA analysis in maternal plasma o If RhIg is not administered, the risk of immunization is only
● Intravascular intrauterine transfusion about 16% for an Rh-negative mother after an Rh-positive
pregnancy
ETIOLOGY__________________________ ● Immunoglobulin Class
o Of the immunoglobulin classes, only IgG is transported across
● Levine, Stetson – reported a transfusion reaction from transfusing a
the placenta
husband’s blood to a postpartum woman; postulated that the mother
o IgG1 and IgG3 are more efficient in RBC hemolysis than are
had been immunized to the father’s antigen through fetomaternal
IgG2 and IgG4
hemorrhage
● Antibody Specificity
o D antigen – most immunogenic
o C, E, and c – also potent immunogens (associated with ● Noninvasive
moderate to severe cases of HDFN) ● Can reliably predict anemia in the fetus
o Anti – E and anti-c caused severe HDFN ● Color Doppler indicates the direction of blood flow, using red for
o Anti – Kell – considered the most clinically significant in its arterial flow and blue for venous
ability to cause HDFN (of non-Rh system antibodies) ● Measurement is based on the reduced blood viscosity at low
o Anti – Fya, Anti – s, Anti – M, Anti – N, Anti – S, Anti – Jka – hematocrit and the resulting faster velocity
rare cause of HDFN
● Influence of ABO Group CORDOCENTESIS
o When mother is ABO incompatible with fetus = fetomaternal
● Procedure used to obtain a sample of fetal blood
hemorrhage incidence is DECREASED (because of the
● The umbilical vein is visualized at the level of the cord insertion into
hemolysis of ABO-incompatible D-positive fetal RBCs in the
the placenta -> a spinal needle is inserted into the umbilical vein ->
mother’s circulation before the D antigen can be recognized by
a sample of the fetal blood is obtained
her immune system)
● Fetal blood tests: (1) hemoglobin, (2) hematocrit, (3) bilirubin, (4)
blood type, (5) direct antiglobulin test, and (6) antigen phenotype
PATHOGENESIS_____________________ and genotype
HEMOLYSIS, ANEMIA, AND ERYTHROPOIESIS AMNIOCENTESIS

● Hemolysis occurs when maternal IgG attaches to specific antigens ● For management of HDFN; used to estimate the extent of fetal
of the fetal RBCs (the antibody – coated cells are removed from the hemolysis
circulation by the macrophages of the spleen) ● Can be done as early as 10 – 12 weeks
● Erythroblastosis fetalis – term used to describe the stimulation of ● Concentration of bilirubin in the amniotic fluid correlates with the
fetal bone marrow to produce RBCs at an accelerated rate due to degree of fetal anemia; the fluid is subjected to spectrophotometry,
destruction of fetal RBCs (erythroblasts are released into the so AOD (optical density) at 450 nm can be calculated
circulation) ● Measurement is plotted on the Liley graph according to gestational
● When the bone marrow fails to produce enough RBCs to keep up age
with the rate of RBC destruction, erythropoiesis is increased in the ● Liley Graph:
hematopoietic tissues of the spleen and liver -> hepatosplenomegaly o Zone I – predict mild or no disease, which do not require
-> portal hypertension and hepatocellular damage intervention
● Hydrops fetalis – the development of high-output cardiac failure o Zone II – most fetuses have moderate diseases that may require
with generalized edema, effusions, and ascites caused by severe intervention
anemia and hypoproteinemias (severe cases – develop by 18 to 20 o Zone III – indicate severe and often life-threatening; hemolysis
weeks) and require urgent intervention
BILIRUBIN INTRAUTERINE TRANSFUSION

● RBC destruction -> releases hemoglobin -> metabolize into bilirubin ● Necessary when one or more of the conditions exists:
(indirect bilirubin) -> indirect bilirubin transported to placenta -> o 1. MCA-PSV indicates anemia
conjugated by maternal liver -> excreted by mother o 2. Fetal hydrops is noted on ultrasound examination
● After birth, the newborn liver is unable to conjugate bilirubin o 3. Cordocentesis blood sample has hemoglobin levels less than
efficiently -> bilirubin can reach levels toxic to the infant’s brain 10 g/dL
(more than 18 to 20 mg/dL) -> if left untreated can cause kernicterus o 4. Amniotic fluid AOD 450 nm results are high
▪ Done by injecting donor RBCs directly into the umbilical
vein; after 36 weeks’ gestation; repeated every 2 to 4
DIAGNOSIS AND MANAGEMENT______ weeks until delivery
▪ Goal: maintain fetal hemoglobin above 10 g/dL
SEROLOGIC TESTING OF THE MOTHER
▪ The hematocrit level of the RBCs to be transfused should
● 1. ABO, Rh, and Antibody Screen – during first prenatal visit, be greater than 70%
preferably during the first trimester
o If the antibody screen is nonreactive, repeat testing is PHOTOTHERAPY
recommended before RhIg therapy
● Phototherapy at 460 to 490 nm is used to change the unconjugated
● 2. Antibody identification – if the antibody is reactive, the antibody
bilirubin to isomers, which are less lipophilic and less toxic to the
identity must be determined
brain
o Sulfhydryl reagent (dithiothreitol or 2 – mercaptoethan) – use
to establish immunoglobukin class
INTRAVENOUS IMMUNE GLOBULIN
● 3. Paternal Phenotype and Genotype – a specimen of the father’s
blood should be tested for the presence and zygosity of the ● Used to treat hyperbilirubinemia of the newborn caused by HDFN
corresponding antigen ● The IVIG competes with the mother’s antibodies for the FC
● 4. Fetal DNA Testing – during second trimester maternal plasma can receptors on the macrophages in the infant’s spleen, reducing the
be tested for fetal DNA to determine genotype amount of hemolysis
● 5. Antibody Titers – the patient serum or plasma is serially diluted
and tested against appropriate RBCs to determine the highest EXCHANGE TRANSFUSION
dilution at which a reaction occurs (titer)
● The use of whole blood or equivalent to replace the neonate’s
o Difference of greater than 2 dilutions, or a score change of more
circulating blood
than 10 – considered a significant change in titer
● Remove high-levels of unconjugated bilirubin; prevents kernicterus
● Other advantages: removal of part of the circulating maternal
COLOR DOPPLER MIDDLE CEREBRAL ARTERY PEAK
antibody, removal of sensitized RBCs, and replacement of
SYSTOLIC VELOCITY
incompatible RBCs with compatible RBCs
● 16 to 20 weeks’ gestation ● Full-term newborn infants = Hb of 14 – 20 g/dL
o Below 10 g/dL – anemia and requires transfusion o After 2000 cells counted
o Below 7 g/dL – severe anemia o Volume of Fetomaternal Hemorrhage:
▪ % of fetal cells x maternal blood volume
SEROLOGIC TESTING FOR THE NEWBORN INFANT # of maternal cells
▪ Then, volume of Fetomaternal Hemorrhage/30 = number
● 1. ABO Grouping – weak reactions are expected due to undeveloped
of required vials of RhIg
ABO Ag in newborns; reverse grouping cannot be used
● 2. Rh Typing – Red cells heavily sensitized w/ anti-D can give a
false (-) Rh type (blocked Rh); an eluate will reveal anti-D, eluted
red cells will show reaction with anti-D
o If DAT is strongly positive, Rh reagents with high-protein
media can also give false (+) results, thus saline reagents are
recommended with a positive DAT
● 3. DAT – most important serologic test for diagnosing HDFN
o Positive DAT: indicates that the antibody is coating the infant’s
RBC
o Strength of the reaction does not correlate well with the severity
of HDN
● 4. Elution – routine preparation is unnecessary; helpful only when
the cause of HDN is in question (resolution of a case of “blocked
Rh”)_
NEWBORN TRANSFUSIONS
● Small aliquot transfusions
● Hemoglobin level of less than 10 g/dL requires transfusion
● Most centers treating HDFN use group O RBCs for intrauterine and
neonatal transfusions
● Rh-negative units: for fetuses and neonates whose blood types are
unknown or are Rh-negative blood transfused to the fetus and
premature infant should also be irradiated to prevent graft-versus-
host-disease
● Blood units should be less than 7 days from collection
MECHANISM OF ACTION OF RhIg____

● Attaches to the fetal Rh-positive RBCs in the maternal circulation
● Antibody-coated RBCs are trapped in the maternal spleen, where
they take up more antibodies form the circulating plasma
● This activates suppressor cell or causes the production of “blocking
ABO – HDN__________________________
Ab” or both ● Maternal ABO Ab that are IgG can cross the placenta and attach to
● Indications ABO-incompatible Ag manifested by hyperbilirubinemia and
o Antenatal: early in the third trimester, or at about 28 weeks’ jaundice within 12 – 48 hours of birth
gestation ● Group O individuals have high-titered IgG Ab than in group A or B,
o Postpartum: Rh-negative nonimmunized mother should receive thus, ABO-HDN is nearly always limited to A or B infants of Group
RhIg soon after delivery of an Rh-positive infant (within 72 hrs O mothers with potent anti-A, B
after birth) ● MILD COURSE of ABO-HDN is due to slow development of ABO
o Other indications for RhIg Ag on fetal RBCs microspherocytes and increased RBC fragility are
▪ Amniocentesis unique characteristics of ABO-HDN
▪ Chorionic villus sampling
▪ Abortion (spontaneous and induced) COMPARISON OF ABO VS RH HDFN___
▪ Ectopic pregnancy Rh ABO
▪ Abdominal trauma First pregnancy Rare Yes
▪ Greater than 40 weeks’ gestation Disease predicted by titers Yes No
▪ Accidental or inadvertent transfusion Antibody IgG Yes (anti-d, etc.) Yes (anti-a, b)
● US regular dose vial (300 ug) – protects against 15 mL of packed Bilirubin at birth Elevated Normal range
RBCs or 30 mL of whole blood Anemia at birth Yes No
● Massive fetomaternal hemorrhages of more than 30 mL of whole
Phototherapy Yes Yes
blood occur in less than 1% of deliveries -> immunization in
Exchange transfusion Common Rare
maternal circulation
Intrauterine transfusion Sometimes None
● A maternal sample should be obtained within 1 hour of delivery and
screened – using a test such as the rosette technique – for massive Spherocytosis Rare Yes
fetomaternal hemorrhage
● If there is massive fetomaternal hemorrhage that is greater than 30 AUTOIMMUNE HEMOLYTIC
mL or positive in rosette technique, quantitation of hemorrhage must
be done through KLEIHAUER – BETKE METHOD ANEMIAS (AIHA)
● Kleihauer – Betke – test to quantitate actual amount of fetomaternal ● Immune hemolytic anemia is defined as shortened RBC survival
hemorrhage; maternal blood smear is treated with acid or alkali and mediated through the immune response (antibodies)
counterstain ● Categories: (1) Alloimmune, (2) Autoimmune, and (3) Drug-
● 1. Fetal cells contain Hb and in which remain pink (acid resistant); induced
maternal cells will appear as ghosts
● Alloimmune response – the patient produces antibodies to foreign or ●Diagnostic tests include DAT and characterization of the
non-self RBC antigens introduced into the circulation through autoantibody in the serum or eluate
transfusion, transplant, or pregnancy ● DAT Panel in AIHA
● Autoimmune Hemolytic Anemia – the autoantibody is directed TYPE OF AIHA ANTI - IgG ANTI – C3d
against the patient’s own RBC; there is a consistent source of WAIHA (67%) + +
antibody and antigen = continuous RBC destruction WAIHA (20%) + -
● Classification of AIHA: (1) Cold reactive (18%), (2) Warm reactive WAIHA (13%) - +
(70%) and (3) Drug-induced COLD
HEMAGGLUTININ
WARM AIHA AND COLD AIHA___________ DISEASE (CHD)
FACTORS WARM AIHA COLD AIHA PAROXYSMAL
COLD
Antibody classification IgG IgM
HEMOGLOBINURIA
Optimal reaction temperature > 32 degrees Celsius < 30 degrees Celsius
(PCH)
Site of hemolysis Usually, extravascular Extravascular/Intravascular
MIXED – TYPE + +
Complement activation May bind complement Binds complement
AIHA (COLD AND
Frequency 70 – 75% of cases 16% of cases WARM)
Specificity Rh antibody Anti I
Reference:
● 1. Warm Autoimmune Hemolytic Anemia (WAIHA) – autoantibody
reacts with patient’s RBC in vivo Notes from Legend Review Center and Micah Vell Diaz, RMT
● 2. Cold Agglutinin Syndrome (CAS) – cold reactive IgM
autoagglutinins binds to RBCs in peripheral circulation (32 degrees Disclaimer: All notes in this material are from the following reference
Celsius) -> IgM binds complement as RBCs return to warmer parts above. No additional notes were included for the creation of this material.
of circulation -> IgM dissociates leaving RBCs coated only with
complement
● 3. Paroxysmal Cold Hemoglobinuria (PCH) – IgG autoantibody
(Anti-P) reacts with RBCs in colder parts of the body, causes
complement to bound irreversibly to RBCs, and cause hemolysis at
warmer temperature
● Drug – Induced
MECHANISM DESCRIPTION IMMUNOGLOBULIN ON
RBC
Drug – Soluble drug – antidrug C3
Dependent or complex Occasionally IgG and IgM
Immune nonspecifically adsorbs
complex loosely to the RBC
(“Innocent surface
Bystander”)
Mechanism
Drug adsorption Drugs such as penicillin IgG
(hapten) or cefotetan bind firmly Rarely C3 may be present
to proteins of the RBC
membrane
Drug – The drug (e.g. alpha - IgG
independent methyldopa/Adomet)
induces production of
an autoantibody that
recognizes RBC
antigen
- Both the
autoantibody and
eluate are reactive
with normal RBCs
Membrane Drugs such as IgG, IgM, IgA. C3
modification cephalosporins modify
the RBCs so that
plasma proteins can
bind to the membrane
nonimmunologically

Review Notes on Blood Banking

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Review Notes on Blood Banking

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BLOOD BANKING

Notes Compiled by: Renz Louie Galanto

• Historical Perspective • Centrifugation

REVIEW NOTES ON BLOOD BANKING

ALLELE_____________________________ DOSAGE EFFECT____________________

STORAGE LESION (LOSS OF RBC VIABILITY)

● Carry oxygen in the absence of RBCs

● Arteriocyte – O-negative prod

BASIC CONCEPTS IN BLOOD BANKING

DOSAGE CHEMICAL REDUCTION OF IGG AND IGM MOLECULES

● Heterozygous alleles has less antigens on red cells (weak

● IgM reacts best at cold temperature (4-22oC) ANTIHUMAN GLOBULIN

● IgM = agglutinates red cells even in 0.85% NSS

ENHANCEMENT MEDIA / POTENTIATORS

● Increases the dielectric constant, which reduces the zeta potential

LOW IONIC STRENGTH SOLUTION MEDIA (LISS)

● Decreases the ionic strength of a reaction medium and so reduce the

4 ALLELE THEORY BY THOMPSON____

● Fresh and have a pH of 7.2 – 7.4 FORMATION OF ABO ANTIGENS

ADDITION OF AHG Gene Glycosyltransferase Immunodominant Acceptor Antigen

Blood Group Antibody Produced Characteristics O GROUP ANTIBODIES

DUFFY ANTIBODIES LUTHERAN BLOOD GROUP SYSTEM

MINOR BLOOD GROUP

OPEN SYSTEM AND CLOSE SYSTEM__

Intermittent – flow centrifugation – one venipuncture site, in

● a. Centrifugation – withdrawal of whole blood, removing selected

CROSSMATCHING___________________ TYPES OF CROSSMATCH

TECHNOLOGIES IN BLOOD BANKING

SOLID PHASE TECHNOLOGY

AFFINITY COLUMN TECHNOLOGY

HEMOLYSIS, ANEMIA, AND ERYTHROPOIESIS AMNIOCENTESIS

BILIRUBIN INTRAUTERINE TRANSFUSION

MECHANISM OF ACTION OF RhIg____

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ALLELE_________ DOSAGE EFFECT