Malaika Ina M.

Galos BSMLS| Adviser: Sir Edgardo Monisit

Immunohematology
READ AT YOUR OWN RISK ▪ transmission of a trait in a predictable fashion from
Pls tell me if nay sayop one generation to the next

WEEK 1 (Lecturer: Sir Edgardo Monisit) LAW OF INDEPENDENT ASSORTMENT

▪ Different pairs of alleles are passed onto the
GENETICS offspring independently of each other. Therefore,
inheritance of genes at one location in a genome
Genetics does not influence the inheritance of genes at
- study of genes, heredity & genetic variation in living another location
organisms ▪ random behavior of genes on separate
- field of biology but intersects frequently with many chromosomes during meiosis that results in a
of life sciences and is strongly linked with the study mixture of genetic material in the offspring
of information system ▪ demonstrates that blood group antigens, inherited
on different chromosome, are expressed separately
Gregor Mendel and discretely
- Father of genetics
- Late 19th century scientist
- Augustinian friar
- Studied trait inheritance patterns in the way traits • Mitosis- occurs in the epithelial cells through binary
were handed down from parents to offspring fission. It is a cell division wherein a parent cell
- Observed that organisms (pea plant) inherit traits by forms two daughter cells with the same number of
way of discrete “units of inheritance” chromosome with the parent cell.
• Meiosis- occurs in the sex cells or gametes. It is a
Gene cell division that results in four daughter cells each
- Region of DNA that encodes a functional “RNA” or with half the number of chromosomes of parent cell
“protein” product, and is the molecular unit of • Endomitosis- occurs in the megakaryocyte. It is a
heredity cell division wherein a parent cell divides without
- Known as unit of inheritance cellular membrane division which results into
- Passed on to an organism’s offspring so that its trait numerous cell copies of chromosomes within the
are inherited parent cell

BASIC TERMINOLOGIES
Genetic loci- sites of a gene on a chromosome
BASIC IMMUNOLOGY IN IMMUNOHEMATOLOGY
Alleles- alternate forms of a gene at a given locus
Codominant- equal expression of two different inherited • Immunology- branch of biology that deals with the
alleles study of the immune system
Recessive- trait expressed only when inherited by both • Antigen- foreign, naturally occuring
parents • Immune system- tightly controlled system in the
Dominant- gene product expressed over another gene body which coordinate to defend a host organism
against intrusion by a foreign substance or abnormal
MENDELIAN LAW OF GENETICS cells or self-origin (tissues, organs, cells & biological
mediators)
• Immunity- refers to the process by which a host
organism protects itself from attacks by external and
LAW OF SEGREGATION internal agents
▪ For any trait, each parent’s pairing of genes split
and one gene (alleles) passes from each parent to an
offspring which particular gene in a pair gets passed
MECHANISM OF THE IMMUNE SYSTEM
on is completely up to chance
▪ passing of one gene from each parent to the Innate or Natural Immunity Adaptive or Acquired Immunity
offspring - Primary line of defense - Suppliment protection
- Nonspecific - Later evolutionary
development
Malaika Ina M. Galos BSMLS- C3 1
 - Natural-present at
births
- Early evolutionary
development

COMPLEMENT PATHWAY

• Classical Pathway- initiated by Ag-ab complexes

INNATE OR NATURAL IMMUNNITY
and requires CIq for activation to proceed
• Alternative Pathway- activated by certain
macromolecules on the cell wall of bacteria, fungi, &
tumor cells & requires C3b, serum factors B, D
properdin & initiating factor
• Lectin Pathway- activated by binding of mannose-
binding-lectin (MBL) to microbe

RBC & PLATELET PRESERVATION

HISTORY

POPE INNOCENT VII (1492)

ANTIBODY STRUCTURE
▪ First transfusion was performed.
▪ blood was taken from three young men and given;
unfortunately, all four died.

BRAXTON HICKS (1869)

▪ recommended sodium phosphate

KARL LANDSTEINER (1901)

▪ discovered the ABO blood groups.
▪ explained the serious reactions that occur in
humans as a result of incompatible transfusions.
▪ His work in the early 20th century won a Nobel
AHG Prize.
- Targets the Fc portion
- Against the portion of Ab
EDWARD E. LINDEMANN
- Forms visible agglutination
▪ carried out vein-to-vein transfusion of blood by
using multiple syringes and a special cannula for
Fab
puncturing the vein through the skin
- Targets the Ag epitopes
▪ the procedure is time-consuming, complicated
required many skilled assistants
• IgG2- least to cross the placenta
• IgG4- least to activate complement
• IgG1&3the bestest haha- best to activate complement UNGER
▪ designed his syringe-valve apparatus
Malaika Ina M. Galos BSMLS-SP1 2
 ▪ transfusions from donor to patient by an unassisted • 1949- established that purified hemoglobin could
physician became practical restore blood volume and deliver oxygen
• 1950s-1960s- platelet transfusion were given freshly
drawn from whole blood volume and deliver oxygen
ALBERT HUSTIN (1914)
• 1970s- platelet have been prepared from whole
▪ reported the use of sodium citrate as an
blood from concentrate in which the volume per
anticoagulant solution for transfusions
unit is near 50mL in contrast to the 250-300 mL
volume of platelet-rich plasma units
RICHARD LEWISOHN (1915) • 1980s- Platelet Additive Solution (PAS) were first
▪ determined the minimum amount of citrate needed developed
for anticoagulation and demonstrated its nontoxicity • 1984- FDA extended platelet storage from 5 to 7
in small amounts days at 20C to 24C
▪ transfusions became more practical and safer for • 1995- PASs have been used in Europe
the patient. • 2002- The use of Nucleic Acid Amplification Tech.
(NAT) licensed by the FDA is one of the reason for
FRANCIS ROUS AND J.R. TURNER (1916) the increased safety of blood transfusion
▪ introduced a citrate dextrose solution for the • 2002- the College of American Pathologists added a
preservation of blood requirement that laboratories have a method to
▪ 1916- glucose was evaluated as preservative screen platelets for bacterial contamination
solutions for the enhancement of RBCs • 2004- AABB introduced a similar requirement
▪ 1930-function of glucose in RBC metabolism was (referring to CAP in 2002)
understood • 2005- FDA approved a study “Post Approval
Surveillance Study of Platelet Outcomes, Released
Tested” (PASSPORT) to collect data on the safety of
WORLD WAR II apheresis platelets tested with FDA-approved
▪ stimulated blood preservation research because the bacterial detection test and stored for 7 days
demand for blood and plasma increased • 2005- the FDA approved the use of prestorage-
▪ Charles Drew pooled platelets prepared by Acrodose PL System
o Developed techniques in blood transfusion and • 2007- The National Blood Collection and Utilization
blood preservation which led to the Survey found that only 175 of platelets doses in U.S.
establishment of a widespread system of blood were whole-blood derived (WBD)
banks • 2008- The Defense Advance Research Projects
o February 1941- he was appointed director of Agency (DARPA) awarded a bioengineering company
the first American Red Cross blood bank at named Arteriocyte contact to develop a system for
Presbyterian Hospital producing O-negative RBCs on the battlefield.
• 2009 (November)- the FDA approved the first rapid
LOUTIT AND MOLLISON (1943) test to detect bacteria in platelets—the Pan Genera
▪ introduced the formula for the preservative acid- Detection (PGD) test (Verax Biomedical)
citrate-dextrose (ACD) • 2011- the time when INTERCEPT platelets were
phased into routine use in Switzerland
• 2014- FDA approved a pathogen reduction
JULY 1947 technology for apheresis platelets stored for up to 5
▪ Issue of the Journal of Clinical Investigation which days as a measure to reduce the risk of bacterial
devoted nearly a dozen papers to the topic of blood contamination of platelets
preservation
• 2016 (March)- recommendation for secondary
▪ blood banks were established in many major cities
testing of platelets prior to issuing
of the United States; subsequently, transfusion
became commonplace

GIBSON (1957)
▪ introduced an improved preservative solution called
citrate-phosphate-dextrose (CPD), which was less
acidic and eventually replaced ACD as the standard
preservative used for blood storage.

Malaika Ina M. Galos BSMLS-SP1 3

WEEK 2 (Lecturer: Doc Apita <3) ▪ Individuals normally produce antibodies directed
against the A and/or B antigen(s) absent from their
RBCs
ABO BLOOD GROUP
▪ These Abs are naturally occurring
▪ are predominantly IgM, activate complement, and
ABO system react at room temperature or colder
- most important of all blood groups in both ▪ production is initiated at birth
transfusion and transplant medicine ▪ m ost antibodies found in cord blood serum are of
- only blood group system in which individuals already maternal origin
have antibodies in their serum to antigens that are ▪ Antibody production peaks between 5 and 10 years
absent from their red blood cells (RBCs) without any of age and declines later in life
prior exposure to RBCs
- incompatible blood transfusion may result in
immediate lysis INHERITANCE OF THE ABO BLOOD GROUPS
- transfusion of wrong ABO group- no.1 cause of ▪ In 1924, ABO inheritance was describe by indicating
death in Hemolytic Transfusion Reaction that an individual inherits one ABO gene frome each
- Transfusion Related Acute Lung Injury- most parent and that these two genes determine which
frequent cause of death (2015) ABO Ags are present on the RBC membrane
▪ inheritance of ABO genes, therefore, follows simple
Mendelian genetics (Mendelian Law of Inheritance)
HISTORICAL PERSPECTIVE & ROUTINE ABO
▪ ABO is codominant in expression
TESTING ▪ Located on chromosome 9
▪ O gene is considered an amorph, as no detectable
Karl Lansdsteiner antigen is produced; O phenotypes is autosomal
- discovered the first human blood group system, ABO recessive
- in 1901, he drew blood from himself & 5 associated, ▪ The designations group A and B refer to phenotypes,
separated cells and serum and mixed each cell whereas AA, BO, and OO denote genotypes
sample w/ each serum ▪ In the case of an O individual, both phenotype and
- first individual to perform reverse & forward genotype are the same because that individual
grouping would have to be homozygous for the O gene. An
individual who has the phenotype A (or B) can have
Forward grouping (front type) the genotype AA or AO (or BB or BO)
- uses known antisera (anti-A, anti-B) to detect ▪ The phenotype and genotype are the same in an AB
antigens on individual’s RBCs individual because of the inheritance of both A and
Reverse grouping (back type) B gene
- detects ABO antibodies in the patient’s serum by
using known reagent RBCs:
o A1 FORMATION OF A, B, AND H RED BLOOD CELL
o B cells ANTIGENS
▪ The formation of ABH antigens results from the
ABO grouping interaction of genes at three separate loci (ABO, Hh,
- Most frequently performed test in the blood bank and Se)
- There is always an inverse reciprocal relationship o ABO, Hh- rbc membrane
between the forward and reverse type o ABO, Se- saliva, tears, digestive juices etc.
o example, if the individual has A antigens
only on their red blood cells, there will be
an “expected” naturally occurring anti-B
antibody in their serum since they lack the
B antigen
- Ab production in most other blood groups requires
the introduction of foreign RBCs by either
transfusion or pregnancy
- Serum grouping is unique to the ABO blood group
system

ABO ANTIBODIES
▪ terminal galactose on the precursor substance is
attached to the N-acetylglucosamine in a beta 1-4
Malaika Ina M. Galos BSMLS-SP1 4
 linkage, the precursor substance on erythrocytes is
referred to as type 2
▪ H antigen is the precursor structure on which A & B
antigens are made
▪ FUT 1 (H) & FUT 2 (Se) genes are closely linked and
located on chromosome 19 & are not part of ABO
system
▪ ABH antigen develop as early as 37th day of fetal life
▪ Expression of A & B antigens on the RBCs is fully
developed by 2 to 4 years of age and remains
constant throughout life

A SUBGROUPS
▪ Group A RBCs that react with both anti-A and anti-
A1 are classified as A1
▪ Cells of approximately 80% of all group A (or AB)
individuals are A1 (or A1B), remaining 20% are A2
(or A2B)

WEAK A SUBGROUPS
▪ Subgroups weaker than A2 occur infrequently and
are most often recognized through an ABO
discrepancy (unexpected reactions in the forward
and reverse grouping)
▪ Characteristics of weak A subgroups include the
following:
INTERACTION OF Hh & ABO GENES o Decreased number of A antigen sites per
▪ Individuals who are blood group O inherit at least RBC (resulting in weak or no agglutination
one FUT 1(H) gene (genotype HH or Hh) and two O with human polyclonal anti-A)
gene o Varying degrees of agglutination by human
▪ H gene elicits the production of an enzyme called anti-A,B8
alpha-2-L-fucosyltransferase that transfers the o Increased variability in the detectability of
sugar L-fucose to an oligosaccharide chain on the H antigen, resulting in strong reactions with
terminal galactose of type 2 chain anti-H
▪ Therefore, L-fucose is the sugar responsible for H o Presence or absence of anti-A1 in the
specificity serum
▪ O blood group has the highest concentration of H
antigen A3 RBCs
▪ term Bombay has been used to refer to the - characteristically demonstrate a mixed-field pattern
phenotype that lacks normal expression of the ABH of agglutination with anti-A and most anti-A,B
antigens because of the inheritance of the hh reagents
genotype o Mixed-field can be defined as small
▪ In the formation of blood group A, the A gene (AA or agglutinates within predominantly
AO) a codes for production of alpha-3-N unagglutinated RBCs.
acetylgalactosaminyltransferase, which transfers an - A3 enzyme is a product of an allele at the ABO locus
N-acetyl-D-galactosamine(GalNAc) sugar to the H inherited in a dominant manner
substance - Anti-A1 may be present in serum of A3 individuals,
▪ Individuals with blood group B inherit a B gene (BB and A substance is detected in the saliva of A3
or BO) that codes for the production of alpha-3-D- secretors
galactosyltransferase and attaches D-galactose
(Gal) sugar to the H substance previously placed on Ax RBCs
the type 2 precursor substance through the action - not agglutinated by anti-A reagent but do
of the H [Link] sugar is responsible for B agglutinate with most examples of anti-A,B
specificity

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 - a transferase is not usually detectable in the serum o Presence or absence of ABO isoagglutinins
or in the RBC membranes of Ax individuals. in the serum
- Ax individuals almost always produce anti-A1 in their o Adsorption-elution studies with anti-B
serum o Presence of B substance in saliva
o Molecular testing
Aend RBCs
- demonstrate very weak mixed-field agglutination B3 RBCs
with some anti-A and anti-A,B reagents - results from the inheritance of a rare gene at the
- No A glycosyltransferase is detectable in the serum ABO locus and is characterized by a mixed-field
or in the RBC membranes of Aend individuals pattern of agglutination with anti-B and anti-A,B
- Aend is inherited as an allele at the ABO locus antisera
- B glycosyltransferase is present in the serum but
Am RBCs not in the RBC membranes of these individuals.
- not agglutinated, or are agglutinated only weakly, - is the most frequent weak B phenotype
by anti-A or anti-A,B reagents.
- An A enzyme of either the A1 or A2 type previously Bx RBCs
described is detectable in the serum of Am - demonstrate weak agglutination with anti-B and
subgroups. anti-A,B antisera
- Am is inherited as a rare allele at the ABO locus - B glycosyltransferase has not been detected

Ay RBCs Bm RBCs
- not agglutinated by anti-A or anti-A,B reagents - unagglutinated by anti-B or anti-A,B antisera
- Trace amounts of A glycosyltransferase is - B glycosyltransferase is present in the serum of
detectable in the serum of Ay individuals, and saliva Bmphenotypes but is usually lower in activity and
secretor studies demonstrate H and A substance, varies from individual to individual
with A substance present in below-normal - Reduced activity of B enzyme in hematopoietic
quantities. tissue is clearly the defect causing the formation of
- This phenotype does not represent expression of an the Bm subgroup
alternate allele at the ABO locus but rather as a
germline mutation of an A gene within a family Bel RBCs
- unagglutinated by anti-B or anti-A,B anti-sera
Ael RBCs - No B glycosyltransferasehas been identified in the
- unagglutinated by anti-A or anti-A,B reagents serum or RBC membrane of Bel individuals
- No detectable A enzyme activity can be - Inherited as a unique mutation in exon 7 of the B
demonstrated in the serum or in the RBC gene at the ABO locus
membranes of Ael individuals
- The Ael phenotype is inherited as a rare gene at the
ABO locus

ABO DISCREPANCIES
→ Occurs when unexpected reactions are observed in
the forward and/or reverse grouping
→ It is important to make sure any and all technical
WEAK B SUBGROUPS
factors that may have given rise to the ABO
▪ Inheritance of B subgroups, similar to that majority
discrepancy are reviewed and corrected
of A subgroups, is considered to be a result of
→ It is also essential to obtain adequate information
alternate alleles at the B locus
regarding the patient’s age, diagnosis, transfusion
▪ Criteria used for differentiation of weak B
history, medications and history of pregnancy
phenotypes include the following techniques:
o Strength and type of agglutination with
anti-B, anti-A,B, and anti-H

Malaika Ina M. Galos BSMLS-SP1 6

 o Leukemias may yield weakened A or B
antigens and Hodgkin’s disease has been
reported in some cases to mimic the
depression of antigens found in leukemia
o The “acquired B” phenomenon will show
weak reaction with anti-B antisera and is
most often associated with diseases of the
digestive tract (e.g., cancer of the colon).

GROUP III DISCREPANCIES

▪ These discrepancies between forward and reverse
groupings are caused by protein or plasma
abnormalities and result in rouleaux formation or
GROUP I DISCREPANCIES pseudoagglutination, attributable to the following:
▪ Group I discrepancies are associated with o Elevated levels of globulin from certain
unexpected reactions in the reverse grouping due to disease states, such as multiple myeloma,
weakly reacting or missing antibodies Waldenström’s macroglobulinemia, other
▪ Common populations with discrepancies in this plasma cell dyscrasias, and certain
group are: moderately advanced cases of Hodgkin’s
o Newborns (ABO antibody production is not lymphomas
detectable until 3 to 6 months of age) o Elevated levels of fibrinogen
o Elderly patients (production of ABO o Plasma expanders, such as dextran and
antibodies is depressed) polyvinylpyrrolidone
o Patients with a leukemia (e.g., chronic o Wharton’s jelly in cord blood samples
lymphocytic leukemia) or lymphoma (e.g.,
malignant lymphoma) demonstrating
hypogammaglobulinemia GROUP IV DSICREPANCIES
o Patients using immunosuppressive drugs ▪ These discrepancies between forward and reverse
that yield hypogammaglobulinemia groupings are due to miscellaneous problems that
o Patients with congenital or acquired have the following causes:
agammaglobulinemia or immunodeficiency o Cold reactive autoantibodies in which RBCs
diseases are so heavily coated with antibody that
o Patients with bone marrow or they spontaneously agglutinate,
hematopoietic progenitor stem cell (HPC) independent of the specificity of the
transplants (patients develop reagent antibody
hypogammaglobulinemia from therapy and o Circulating RBCs of more than one ABO
start producing a different RBC population group due to RBC transfusion or
from that of the transplanted bone marrow/stem cell transplant
marrow) o Unexpected ABO isoagglutinins
o Patients whose existing ABO antibodies o Unexpected non-ABO alloantibodies
may have been diluted by plasma
transfusion or exchange transfusion
o ABO subgroups

GROUP II DISCREPANCIES

▪ Group II discrepancies are associated with

unexpected reactions in the forward grouping due
to weakly reacting or missing antigens
▪ This group of discrepancies is probably the least
frequently encountered
▪ The following are some of the causes of
discrepancies in this group:
o Subgroups of A or B may be present

Malaika Ina M. Galos BSMLS-SP1 7

WEEK 3 (Lecturer: Sir Edgardo Monisit) ▪ an individual’s Rh phenotype is reported as DCE
instead of CDE because Fisher-Race postulated that
the C/c locus lies between D/d and E/e loci
RH BLOOD GROUP
▪ haplotype- combination of maternal and paternal
genes inherited (ex. DCe/Dce)
HISTORY ▪ there are rare phenotypes that involve deletions of
specific genes, and in those cases the deletion is
Levin and Stetson (1939) represented with a dash (ex. D-)
- Described an HTR in obstetrical patient ▪ placing parenthesis around (D), (C), and (e) indicates
- After delivering a stillborn infant, the woman weakened antigen expression
required transfusions. Her husband, who had the
same ABO type, was selected as her donor. After
transfusion, the recipient demonstrated classic
symptoms of an acute hemolytic transfusion
reaction (AHTR)
- HDFN ins manifested in the 2nd exposure of
pregnancy and Rh positive baby
- Antibody was detected in the mother’s serum which
reacted in both 37oC (IgG) and 20oC (IgM)
- They postulated that the fetus and the father
possessed a common factor which was not
- Levine and co-workers – same blood group yet
different Ag. – weiner:RH-HR terminology—
▪
Landsteiner Weiner
- Described an antibody made by guinea pigs and
rabbits when they were transfused with rhesus WIENER-RH-Hr TERMINOLOGY
macaque monkey RBCs ▪ Wiener believed there was one gene responsible for
o The antibody agglutinated 85% of human defining Rh that produced an agglutinogen
RBCs and was named anti-Rh after the containing 3 Rh factors
rhesus monkey ▪ the original Wiener nomenclature named the five
common Rh antigens as Rho, rh’, rh’’, hr’, and hr’’,
• Rh- human-produced antibody ▪ an agglutinogen is described by a letter and symbol
• Anti-LW- anti-rhesus formed by the animals assigned based on the factors present
• 1940s-five antigens were defined in the Rh system o R - presence of D Ag
• 1980s- molecular testing further defined the structure of o r- absence of D Ag
RH genes o 1 or ‘- presence of C Ag
TERMINOLOGY o no 1 or ‘ - presence of c Ag
o 2 or ‘’- presence of E Ag
→ There are four terminologies used to describe the o no 2 or ‘’- presence of e Ag
Rh system o z or y – both CE are present
→ Phenotype is the serologic detection of antigens o
using specific antiseraa Antigens Blood factors Modified designation
→ Genotype is an individual’s genetic makeup D Rho R
d r
FISCHER-RACE: DCE TERMINOLOGY C rh’ 1 or ‘
▪ In 1940s Fischer and Race defined the five common c rh” no 1 or
Rh antigens E hr’ 2 or ‘’
o (D/d, C/c, and E/e) e hr” no 2 or ‘’
▪ an individual inherits a set of RH genes from each
parent (i.e., one D or d, one C or c, and one E or e)
▪ Postulated that Antigens of Rh system were ▪ The genotype for the Rhnull that arises from an
produced by the three closely linked sets of alleles amorphic gene at both Rh loci is written as rr(w/ ⁼ sa
▪ Each gene produces a product on the RBC surface upper part) and pronounced “little r double bar”
▪ Each antigen and the corresponding gene is assigned ▪ This terminology allows one to convey Rh antigens
w/ the same letter inherited on one chromosome or haplotype and
▪ D Ag- non immunogenic; most exposed makes it easier to discuss a genotype

Malaika Ina M. Galos BSMLS-SP1 8

 ▪ Note:an agglutinogen in Wiener nomenclature ▪ Universal language of blood bankers
actually represents the presence of a single ▪ Its mandate was to establish a uniform
haplotype expressing three different antigens nomenclature that is both eye- and machine-
▪ In this terminology, there is no designation for the readable and is in keeping with the genetic basis of
absence of D antigen blood groups
▪ adopted a six-digit number for each authenticated
antigen belonging to a blood group system
▪ therefore, D is RH1, C is RH2, and so forth
▪ a minus sign preceding the number indicates that
the antigen was tested for but was not present
o the phenotype D + C – E + c + e + or DcE/ce
or R2r would be written RH:1, –2, 3, 4, 5
▪ when referring to a gene, an allele, or a haplotype,
the symbols are italicized (NOTE: A haplotype is
Caucasians: R1 > r > R2 > R0 followed by a space or asterisk)
African-Americans: R0 > r > R1 > R2
GENES
ROSENFIELD & COWORKERS: ALPHANUMERIC
TERMINOLOGY
▪ In 1960s, Rosenfield and associates proposed a RH GENES
system that assigned a number to each antigen of ▪ RHD & RHCE are two closely linked genes located on
the Rh system in order of its discovery or recognized chromosome 1 that control expression of Rh
relationship to the Rh system proteins
▪ Each antigen is assigned a number. A minus sign ▪ They are codominant
preceding a number designates the absence of the ▪ RHD- codes for the presence of RhD protein
antigen ▪ RHCE- codes for either RhCe, RhcE, Rhce, RhCE
o D is assigned Rh1, C is Rh2, E is Rh3, c is proteins
Rh4, and e is Rh5
RH-ASSOCIATED GLYCOPROTEIN (RHAG)
▪ Resides in chromosome 6 and its product is Rh-
associated glycoprotein
▪ Has a very similar structure to the Rh proteins with
the difference that it is glycosylated which menas
that carbohydrates are attached
▪ It forms complexes with the Rh proteins
▪ Termed as a coexpressor
▪ Must be present for successful expression of Rh
antigens
▪ Does not express any Rh antigens

RH-POSITIVE PHENOTYPES
▪ Inherited as codominant alleles
▪ Inherit one or two RHD genes

RH-NEGATIVE PHENOTYPES
▪ The RBCs lack detectable D antigen
▪ Arise from several pathways
▪ The most common Rh-neg results from the
complete deletion of RHD gene (individuals possess
no RHD gene but have inherited two RHCE genes)

MOST PROBABLE GENOTYPES

▪ Determining most probable genotypes was used in
the past for parentage studies, also known as
INTERNATIOL SOCIETY OF BLOOD TRANSFUSION relationship testing, and for population studies
COMMITTEE: UPDATED NUMERIC TERMINOLOGY
Malaika Ina M. Galos BSMLS-SP1 9
RH ANTIGENS ▪ Anti-D was the most common cause of HDFN until
RH-immune globulin was introduced

BIOCHEMISTRY OTHER COMMON RH ANTIGENS (C,E, c,e)

▪ Nonglycosylated proteins- product of RH genes
▪ C, c, E and e antigens are inherited through the
▪ Rh Ag reside on transmembrane proteins and are
RHCE gene
an integral part of of RBC membrane
▪ Amino acid position 103 is important in determining
C or c expression
▪ Position 226 differentiates E from e
▪ RhD and RhCE proteins and RHAG are exclusively WEAK D: VARIATIONS OF D ANTIGENS
on red blood cells → Serologic weak D is noted when initial anti-D testing
▪ The greatest number of D antigen sites are on cells is negative, or less than or equal to 2+ strong, but
of the rare Rh phenotype D– (refer to the detectable at the indirect antiglobulin testing (IAT)
“Deletions” section) phase
▪ The commonly encountered Rh genotypes, R2R2 cells → Individuals with RBCs carrying weaker D antigen
possess the largest number of D antigen sites (historicallly called Du type) can produce anti-D if
they are missing epitopes of the D antigen
→ Can be separated into three categories:
o Position effect
o Quantitative
o Partial-D antigen

POSITION EFFECT: C IN TRANS TO D

▪ C trans is also known as the position effect or gene
interaction effect
▪ The allele carrying RHD is trans (or in the opposite
haplotype) to the allele carrying C (ex. Dce/dCe)
o Cis- same position
o Trans-opposite position
▪ NOTE: It is not possible to serologically distinguish
ANTIGEN CHARACTERISTICS genetic weak D from the position effect weak D
▪ Rh antigen are proteins integral to the RBC
membrane
▪ Passes through the RBC wall 12 times WEAK D: QUANTITATIVE CHANGES DUE TO
▪ Developed at birth which may cause hemolytic FEWER D ANTIGEN SITES
disease of the fetus and newborn (HDFN) ▪ Complete but fewer in number
▪ mutations in the RHD gene occur, causing changes
or deletions in amino acids present in the
transmembrane or intracellular region of the RhD
protein
▪ activated by mutation of polypeptide
▪ requires IAT for confirmation
▪ Wagner, Gassner, and others described the
classification of weak D RBCS based on single
nucleotide polymorphisms (SNP)
o Mutations for these weak D types occur
because SNP affects amino acid insertion of
the protein on the RBC membrane. When
these changes occur, normal RhD antigen
expression is altered because there are
D ANTIGEN fewer antigen sites overall.
▪ Highly immunogenic and is the most potent
▪ NOTE: exposure to less than 0.1 mL of Rh -positive
RBCs can stimulate Ab production in an Rh-negative D el
person ▪ phenotype occurring in individuals whose red blood
▪ 85%- Rh+ : 15%-Rh- ---in general population cells possess an extremely low number of D antigen
sites that most reagent anti-D are unable to detect

Malaika Ina M. Galos BSMLS-SP1 10

 ▪ Molecular studies can detect mutant RHA genes that ▪ IgG3 and IgG1 – best in fixing complement
alters the expression of RhD patients ▪ IgG2 and IgG4 – least in fixing complement
▪ adsorbing and eluting is oftern the only way to ▪ Coating antigen for (enhance?) phagocytes
detect the D antigen (opsonization)
▪ common in Asians; rare in white ▪ Neutralizes toxins and viruses(?)
▪ Participates in agglutination and precipitation
reactions
PARTIAL D / D MOSAIC
▪ IgM Rh antibodies are formed initially, followed by a
▪ when one or more D epitopes within the entire D
transition to IgG
protein is either missing or altered
▪ An individual with low-titer Rh antibody may
▪ partial-D antigens may type weaker than expected
experience an anamnestic (secondary) antibody
or that may not react at all when routine procedures
response if exposed to the same sensitizing antigen
are used with most commercial anti-D reagents
▪ They do not bind complement
o due to mutation of RHD gene
▪ RBC destruction resulting from Rh antibodies is
o common in individual in asian ethnicity
primarily extravascular

D EPITOPES ON RHCE PROTEIN

RH ANTIBODIES IN PREGNANCY
▪ RhCE protein can express RhD epitopes detected by
▪ Rh Abs formed by pregnant women cross the
some monoclonal anti-D
placenta and may coat fetal RBCs that carry the
▪ There are rare individuals who possess these
corresponding antigen
unusual proteins and when typed with anti-D will
o Which results in fetal cells having positive
show positive reactivity even though the D epitope
(+) DAT
is on the RhCE protein
o Ex. DHAR and ceCF
o R0 Har (RH33) results from a hybrid gene RH ANTIBODY FORMATION
RHCE-RHD-RHCE, in which only a small - IgM – IgG
portion of RHD is inserted into the RHCE - Do not bind complement
gene - 120 days – 1st exposure
- 2-3days – 2nd exposure

Weak D issues
- Typed as D positive but sometimes patients who TESTING FOR RH ANTIGENS & ANTIBODIES
received transfusion from D positive individuals
produce anti-D, this is due to the absence of some of
the antigenic substances present in transfused D RH TYPING REAGENTS
positive individuals ▪ Reagents used to type for D and for the other Rh
Determination of D status antigens may be derived from a variety of sources
- When a weak D individual is the donor he/she must o high-protein based or low-protein based
be typed Rh(D) positive o saline based
- When a weak D is the patient he/she must be typed o chemically modified
Rh(D) negative o monoclonal
- Note: this is to avoid alloimmunization o blends of monoclonals

RH ANTIBODIES

CHARACTERISTICS OF RH ANTIBODIES
▪ most are IgG & react at 37C
▪ produced following exposure of the individual’s
immune system to foreign RBCs
▪ enhanced when testing w/ enzyme treated RBCs
▪ IgG1, IgG2, IgG3, and IgG4 are subclasses of Rh
antibodies
▪ IgG1 & IgG3 – greatest clinical significance because
the reticuloendothelial system rapidly clears RBCs
coated with IgG1 & IgG3
▪ IgG2 – producing immunity for newborn(least)
Malaika Ina M. Galos BSMLS-SP1 11
CLINICAL CONSIDERATIONS
→ Rh antigens are highly immunogenic
→ D antigens most immunogenic outside of ABO
HDN
- Cause of death is severe anemia
- RhIG (intervention)
o 20mg/dl – normal level of unconjugated
bilirubin
RH DEFICIENCY SYNDROME

Rhnull
→ demonstrate a mild compensated hemolytic anemia,
reticulocytosis, stomatocytosis, a slight-to-moderate
decrease in hemoglobin and hematocrit levels, an
increase in hemoglobin F, a decrease in serum
haptoglobin, and possibly an elevated bilirubin level

Regulator
- A mutation occurs in the RHAG gene
- no RhAG protein expression and subsequently no
RhD or RhCE protein expression on the RBCs
- these individuals can pass normal RHD and RHCE
genes to their children
Amorphic
- there is a mutation in each of the RHCE genes
inherited from each parent as well as the deletion of
the RHD gene
Rhmod
→ exhibit RBC abnormalities similar to Rhnull
syndrome; however, clinical symptoms are usually
less severe & rarely clinically remarkable

UNUSUAL TYPES AND RARE ALLELES

Cw
f (ce)- when c & e are joined together(cis)
rhi (Ce)- when C & e are joined together
G- present on most D-positive and all C-positive RBCs
Rh17 (Hr0)
Rh23, Rh30, Rh40, and Rh52
Rh33 (Har)
Rh32
Rh43 (Crawford)
e Variants
V and VS-
Deletions

Malaika Ina M. Galos BSMLS-SP1 12

WEEK 4 (Lecturer: Sir Edgardo Monisit) → If the mother and fetus are ABO incompatible, the
incidence of detectable fetal-maternal hemorrhage
HEMOLYTIC DISEASE OF THE FETUS & is decreased
NEWBORN
• also known as “erythroblastosis fetalis” PATHOGENESIS OF HDN
• alloimmune condition that develops in fetus → Hemolysis occurs when maternal IgG antibody
• IgG Abs from the mother crosses the placenta & attaches to specific antigens of the fetal RBCs
destroys the fetal red blood cells which contains the → Antibody coated cells are removed from the
specific antigen circulation by the macrophages
• Could cause fetal death (hydrops fetalis) → Erythropoietin is released due to anemia and
stimulates the bone marrow to produce RBCs in an
Clinical Signs & Symptoms increased rate
- Jaundice (increased fetal bilirubin level that could
cause “kernicterus”) DIAGNOSIS & MANAGEMENT
- Acute anemia
- Respiratory distress A. Serologic Testing
- Splenomegaly - Type and AB screen- done during the first prenatal visit
o Increased RBC destruction preferrably during the 1st trimester
o Increased activity of the spleen B. ABO & Rh Testing
o Spleen- the graveyard of RBC - the mother’s RBCs must be tested with ABO & Rh, and if
- Hepatomegaly no Rh reaction has occured weak D typing may be
induced
Blood Groups that causes HDN C. Antibody Screening
1. ABO- common but mild - must be able to detect clinically significant IgG
2. Rh- severe alloantibodies that are reactive at 37°C and in the
3. Kell antiglobulin phase
4. Kidd - atleast two separate reagent screening RBCs that
5. Lewis express all of the common blood group antigens
6. Duffy (preferably homozygous) should be used
7. MN - an antibody-enhancing medium such as polyethylene
8. P glycol (PEG) or low ionic strength solution (LISS) can
increase sensitivity of the assay
ABO HDN D. Antibody Identification
- Cold reactive IgM antibodies such as anti-I,
→ Occurs in women with blood type O with an
anti-IH, anti-Lea, anti-Leb, and anti-P1 can be ignored
A(white)/B(black) fetus
-anti-D, the most common and most significant antibodies
→ Condition rarely happens because predominantly
are anti-K, anti-E, anti-c, anti-C, and anti-Fya
ABO Abs are of IgM type
- used in order to assess what particular Ab is present in
→ Abs responsible is the anti-A, anti-B, anti-A,B, IgG
the maternal serum
Abs
E. Paternal Phenotyping
→ Happens in 3% of the total population
- presence of zygosity of a corresponding Ag (Dce/dcE)
→ Fetal hyperbilirubinemia jaundice will manifest (AO, BO, AA, BB, AB)
within 12 to 48 hours after birth and can be treated
F. Amniocyte Testing
with phototherapy - done as early as 10-12 weeks gestation for the
→ severe cases will require exchange transfusion but determination of the D Ag carried by amniocytes
extremely rare G. Antibody Titer
→ mother & infant are ABO incompatible in 1 of every - The relative concentration of Abs that could cross the
5 pregnancies placenta & cause HDN must be determined by antibody
titration.
RH HDN - Note: it is important because the higher the titer, the
→ caused by the sensitization of an RH negative more intense the action of the Ab against the fetal RBCs
mother with an RH positive fetal blood during fetal- H. Amniocentesis & Cordiocentesis
maternal hemorrhage during delivery abortion and - Amniocentesis is done to assess the status of the fetus.
miscarriage The concentration of bilirubin in the amniotic fluid
→ As little as 1 mL of fetal RBCs can immunize the correlates to the severity of fetal anemia.
mother - Lecithin:Sphingomyelin ratio- detection of fetal lung
maturity
Malaika Ina M. Galos BSMLS-SP1 13
 - Cordocentesis- RBCs are injected into the peritoneal → An active immunization induced by RBC Ag can be
cavity..... (pic from PPT) prevented by the administration of the
- Cordocentesis or the percuataneous umbilical cord corresponding RBC antibody (passive immunization)
sampling is used to collect blood from the umbilical vein o RhIG is a concentrate of IgG anti-D
and tested for hemoglobin level..... (23G needles) prepared from pools of human plasma
I. Intrauterine Transfusion → This principle has been used to prevent
- is performed by accessing the fetal umbilical vein immunization of the Rh negative to the Rh positive
(cordocentesis) and injecting donor RBCs directly into the fetal cells by the use of high-tittered RhIG
vein. The goal of intrauterine transfusion is to maintain → Note: During the administration of RhIG there’s an
fetal hemoglobin above 10 g/dL. Once intrauterine immune system blockage. Mother’s immune system
transfusion is initiated, the procedure is typically repeated is blocked by the RhIG through binding to the RhD
every 2 to 4 weeks until delivery to suppress fetal from the fetal red blood cells thus causing the lysis of
hematopoiesis. The initial intrauterine transfusion is those cells. Once lysed, it…..
rarely performed after 36 weeks’ gestation.
- performed when:
• MCA-PSV indicates anemia (>1.5 MoM) MECHANISM OF ACTION
• Fetal hydrops is noted on ultrasound examination ▪ RhIG attached to the fetal Rh positive RBCs in the
maternal circulation. The amount of necessary
• Cordocentesis blood sample has hemoglobin level
antibody to prevent alloimmunization has been
less than 10 g/dL
determined experimentally and is known.
• Amniotic fluid ΔOD 450 nm results are high and/or
increasing.
INDICATION OF RHIG
J. Early Delivery Antenatal
- used to interrupt the transport of maternal antibody to ▪ RhIG must be given early in the 3rd trimester or at
the fetus and to allow exchange transfusion about 28 weeks gestation
K. Phototherapy Post Partum
- converts bilirubin to biliverdin ▪ Rh-neg. non-immunized mother should receive RhIG
soon after delivery of an Rh-positive infant. The
SEROLOGIC TESTING IN THE NEWBORNS recommended interval is w/in 72 hrs. after delivery

A. ABO Grouping DOSE & ADMINISTRATION

- reverse grouping is not used ▪ The regular-dose vial of RhIG in the United States
B. Rh Typing contains sufficient anti-D to protect against 15 mL of
- blocked RH may occur. Elution is done in order to packed RBCs or 30 mL of whole blood
remove anti-D attached to the infant RBCs
C. Direct Antiglobulin Testing
- most important test to diagnose HDN
D. Elution
- done in order to remove IgG CALCULATION OF FETOMATERNAL
HEMORRHAGE
NEWBORN TRANSFUSIONS
𝑛𝑜. 𝑓𝑒𝑡𝑎𝑙 𝑐𝑒𝑙𝑙𝑠 × 𝑚𝑎𝑡𝑒𝑟𝑛𝑎𝑙 𝑏𝑙𝑜𝑜𝑑 𝑠𝑎𝑚𝑝𝑙𝑒
A. Small Aliquot Transfusion 𝑣𝑜𝑙. 𝑜𝑓𝐹𝑀𝐻 =
𝑛𝑜. 𝑜𝑓 𝑚𝑎𝑡𝑒𝑟𝑛𝑎𝑙 𝑐𝑒𝑙𝑙𝑠
- done when there is low bilirubin level in the circulation
and low hemoglobin & hematocrit
Kleihauer betke test
B. Exchange Transfusion
- measure fetal cells vs. Maternal cells
- done when there is high bilirubin level, low
-determine fetal cells in maternal circulation
hemoglobin, low hematocrit in the circulation

SELECTION OF BLOOD FOR TRANSFUSIONS HEMOGLOBIN F STAIN

▪ Acid elution test is employed to assess the
→ Intrauterine transfusion requires donor hematocrit distribution of hemoglobin F in the RBCs. This
→ One practice of exchange transfusion is to prepare information is useful to… . In determination of the
RBCs from whole blood units (Group O RBCs & presence red cells in the maternal circulation during
Group AB Plasma) pregnancy.

RHIG REAGENT & EQUIPMENT

▪ Fetal cell fixing solution is ethyl alcohol, 80%
Malaika Ina M. Galos BSMLS-SP1 14
 ▪ Fetal cell buffer solution is citric-acid phosphate 9. Counter stained the smears with erythrosin B for 4
buffer, pH 3.2 to 3.3 minutes. Rinse with distilled water, allow to air dry, and
▪ Fetal cell stain cover slip (if desired).
1. Erythrosin B (Eosin B) stain, 0.1% 10. Examine the slides microscopically, (oil immersion
2. Ehrlich’s acid hematoxylin objective [1000]).
o Coplin jars
o Water bath, 37C
PROCEDURE
SPECIMEN ▪ To determine the percentage of red blood cells
▪ Blood smears from venous blood collected in EDTA containing fetal hemoglobin:
anticoagulant or from the fingertip (toe or heel) 1. Determine the average number of red cells/field
▪ For best result, blood should be less than 6 hrs. old, 2. 𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝐻𝑔𝑏 𝐹 𝑅𝐵𝐶 𝑝𝑒𝑟 𝑓𝑖𝑒𝑙𝑑 =
𝑛𝑜. 𝑜𝑓 ℎ𝑔𝑏 𝐹 𝑅𝐵𝐶 𝑐𝑜𝑢𝑛𝑡𝑒𝑑
𝑛𝑜.𝑜𝑓 𝑓𝑖𝑒𝑙𝑑𝑠 𝑐𝑜𝑢𝑛𝑡𝑒𝑑
although successful staining has been achieve on 3. % 𝑜𝑓 𝑅𝐵𝐶 𝑤𝑖𝑡ℎ ℎ𝑔𝑏 𝐹 =
𝑛𝑜. 𝑜𝑓 ℎ𝑔𝑏 𝐹 𝑅𝐵𝐶 𝑝𝑒𝑟 𝑓𝑖𝑒𝑙𝑑
× 100
𝑎𝑣𝑒𝑟𝑎𝑔𝑒 𝑛𝑜.𝑜𝑓 𝑅𝐵𝐶 𝑝𝑒𝑟 𝑓𝑖𝑒𝑙𝑑
specimen refrigerated for up to 2 weeks
Note: 1 field= 100 RBCs
▪ Smears should be fixed within 2 hrs. of preparation

PRINCIPLE
▪ Blood smears are fixed with ethyl alcohol and then
incubated in a citric-acid buffer solution in an acid
medium (pH 3.2 to 3.3) , hemoglobin F is resistant
to elution from the red blood cells, while other types
are removed from the red blood cells.
▪ The slides are stained with hematoxylin (stains the
white cell nuclei) and erythrosin B (stains the red
cells)
▪ The smears are then reviewed microscopically to
mine the presence of hemoglobin F, and percentage
of red blood cells containing fetal hemoglobin may
be assessed

PROCEDURE

1. Prewarm citric acid buffer. Place 50 mL of the buffer

solution into a coplin jar and cover. Incubate at 37C for
30 minutes (make certain the level of water in the
incubator is level with or above the level of the buffer in
the coplin jar)
2. Preparation of blood-smears, patient-make several
thin blood smears
3. Allow the blood smears to air dry for at least 10
minutes
4. Fix blood smears (patient and controls) in 80% ethyl
alcohol for 5 minutes
5. Rinse the smears carefully in distilled water and
allowed to air dry
6. Place the dry smears in the prewarmed citric acid
phosphate buffer solution for 5 minutes. At 1 and 3
minutes (of incubation), carefully lift each slide out of
the buffer solution and immediately replace. This action
will provide a gentle stirring of the solution.
7. After 5 minutes, remove the slides from the citric
acid phosphate buffer solution and carefully rinse with
distilled water. Air dry.
8. Stain the dry smears in acid hematoxylin for 3
minutes. Rinse with distilled water and remove as much
of the water as possible from the smears by gently
tapping one end of the slide on an absorbent material
Malaika Ina M. Galos BSMLS-SP1 15
WEEK 5 (Lecturer: Sir Edgardo Monisit) (sialoglycoprotein, SGP). There are 1,000,000 of GPA
per RBC
- Defined by the first and fifth amino acids
OTHER BLOOD GROUPS
- M= Ser-Ser-Thr-Thr-Gly
- N= Leu-Ser-Thr-Thr-Glu
LEWIS (007); FUT3; C-19 - Note: these antigens are well developed at birth
→ Has a coexpressor alike (H Ag) - Enzymes that could easily destroy or remove the M
→ Located on short arm chromosome 19 and N antigens:
o Ficin
→ Incorporated in the RBC membrane
o Papain
→ Codominant
o Bromelin
→ Requires an Se gene for it to be secreted
o Trypsin-less common
o If not inherited phenotype is Le(a+b-)
o Pronase-less common
→ Leb Ag- It is the result of the genetic interaction of
- M and N antigens are not detected in lymphocytes,
the Le & Se gene
monocytes, and granulocytes nor in platelets
→ Found in the:
- GPA and M and N have been detected on renal
o Saliva as glycoproteins capillary endothelium and epithelium
o Plasma as glycolipids
→ Note: GI tract is the primary source of Le glycolipid in S and s antigens
the plasma - Located on a smaller glycoprotein called
Glycoprotein B (GPB) that is very similar to GPA.
Leb formation There are 170,000-240,000 copies of GPB per RBC
- When the Se glycosyltransferases attaches an L- - S and s are differentiated by the amino acids at
fucose to the type 1 precursor forms a type 1H position 29 on GPB.
precursor and the Le glycosyltransferase attaches - S-methionine; s-threonine
another L-fucose - There are about 1.5 times more copies of GPB on
Le(a-b-) S+s– RBCs than on S–s+ RBCs
- Absent in RBCs but Le is present in tissue & - Note: S and s antigens are less easily degraded by
secretions caused by specific mutation enzymes because the antigens are located farther
Leb down the glycoprotein and enzyme-sensitive sites
- Has a receptor of Helicobacter pylori (causative are less accessible.
agent of stomach ulcer) - Enzymes that can easily destroy or remove the S and
s antigens
ANTIBODIES o Ficin
→ Occurs in pregnant women because of the dilution o Papain
effect causing the dissociation of Le Ag in the RBC o Bromelin
→ Not clinically significant because they are o Pronase
neutralized by plasma or serum soluble to these Ags o Chymotrypsin
→ Clinically significant Lewis Abs are considered - Note: The amount of degradation depends on the:
insignificant in transfusion o Strength of the solution
o Length of the treatment
Le(a+b-) nonsecretor- type 1H precursor o Enzyme to cell ratio
Le(a-b+) secretor- type 1H substance (has 1 L-fucose)
ANTIBODIES
MNS (002); GYPA,GYPB; C-4 Anti-M
→ Discovered by Landsteiner & Levine - 50%-80% are IgG or have an IgG component
- Do not bind complement (no lysis), regardless of
→ Antibodies reported are anti-M and anti-N
their immunoglobulin class
→ M and N antigens are antithetical
- Do not react with enzyme treated RBCs
→ Later, S antigen is discovered by Walsh and
- Antigen dosage may affect the reaction
Montgomery and is genetically linked to M and N. Its
- Some are pH and glucose dependent, reacting best
antithetical partner s was discovered in 1951
at pH of 6.5 (pH of blood: 7.35-7.45)
- Note: as long as anti-M does not react at 37C, it is
ANTIGENS not considered clinically significant in transfusion
M and N Antigens Anti-N
- Are found on a well-characterized glycoprotein A - A cold reactive IgM or IgG saline agglutinin
(GPA), the major RBC sialic-acid-rich glycoprotein - Does not bind complement
- Does not react with enzyme treated RBCs
Malaika Ina M. Galos BSMLS-SP1 16
 - Can demonstrate dosage effect - Expression changes during fetal development
- Rarely pH and glucose dependent - Found on fetal RBCs as early as 12 weeks, but
- Not clinically significant if nonreactive at 37C weakens with gestational age
Anti-S and Anti-s - Younger fetuses are more P1+ than older fetuses
- Mostly of IgG class - Poorly expressed at birth and may take up to 7 years
- Reactive at 37C and the antiglobulin test phase for full expression
- A few expresses reactivity at 10C – 22C by saline
indirect antiglobulin test.
ANTIBODIES
- Dosage effect can be exhibited
Anti-P1
- May or may not react with enzyme treated RBCs
- Is a common, naturally occuring IgM antibody in the
- Anti-S and Anti-s are more likely clinically significant
sera of P1- individuals
than Anti-M
- Typically a weak, cold, reactive saline agglutinin
optimally reactive at 4C and not seen in routine
DISEASE ASSOCIATION testing
▪ GPA may serve as a receptor by which certain
pyelonephritogenic strains of Escherichia coli gain
LUKE (LKE) GLOBOSIDE COLLECTION (209)
entry to the urinary tract.
▪ Found in patients with Hodgkin’s Lymphoma
▪ Hence: GPA, M, and N have been detected in the
▪ Patients with Hodgkin’s Lymphoma are divided into
renal capillary endothelium and epithelium
three population:
o 84% is LKE+
P BLOOD GROUP (003); A4GALT; C-22 & o 14% is weakly LKE+
GLOBOSIDE BLOOD GROUP (028); B4GALT1; C-3 o 2% is LKE-
→ Comprised of P, P1, and Pk and antigens and later,
Luke DISEASE ASSOCIATION
→ P1 is assigned to the P Blood Group (003) ▪ Anti-P1 is associated with parasitic infection
→ P is assigned to the Globoside Blood Group (028) (Echinococcus granulosus)
→ P Blood group was introduced in 1927 by ▪ Anti-PP1Pk, and anti-P is associated with early
Landsteiner and Levine abortions
→ Human RBCs are initially grouped ito P+ and P- due ▪ Autoanti-P is associated with Paroxysmal Cold
to the discovery of anti-P Hemoglobinuria
→ In 1951 Levine described anti- Tja (now known as
anti-PP1Pk)
I BLOOD GROUP SYSTEM (027); GCNT; C-6
→ Sanger later showed that anti-Tja is related to the P
blood group because it is common P+ and P- cells → Cold agglutinins in the serum of normal individuals
and was made by a P null inidvidual and in patients with acquired hemolytic anemia
→ Thus the renaming of the original antigens and → Weiner gave a name to this agglutinins, calling its
phenotypes are done: antigen “I” for individuality
o Anti-P became anti-P1
o P+ became P1 ANTIGENS
o P- became P2 I and i antigen
o Pnull became p - High incidence antigen
→ Discovery of Pk antigen in 1959 made the P blood - Expressed in a reciprocal relationship that is
group more complex developmentally regulated
o This antigen is expressed on all the RBCs - Infant RBCs are rich in i antigen, I is almost
except for the very rare p phenotype, but is undetectable
not readily detected unless P is absent
→ Note: The two common phenotypes are P1 and P2
ANTIBODIES
Anti-I
ANTIGENS - A common autoantibody that can be found in
▪ P, and P1, and Pk may be found on RBCs, and WBCs virtually all sera, although testing at 4C with enzyme
▪ P can be found on platelets, epithelial cells, and treated RBCs may be required to detect reactivity
fibroblasts - Detected in serum of many normal healthy
▪ P and Pk can be found in: individuals
o plasma as glycosphingolipids - Not associated with in-vivo RBC lysis
o hydatid cyst fluid as glycoproteins Anti-i
P1 antigen

Malaika Ina M. Galos BSMLS-SP1 17

 - Exhibits strong reactions with cord RBCs and adult i - Usually IgG and reactive in antiglobulin phase, but
RBCs and weaker reactions with adult I RBCs some agglutinate in saline-suspended RBCs
- IgM in nature - About 20% bind complement, but they are seldom
- Reacts best with saline-suspended cells at 4C lytic
- Not commonly found in the serum of healthy Anti-kpa, Anti-Jsa and other low-incidence Kell antigens
individuals - Rarely found due to few exposures to these
antigens
Anti-K, Anti-Kpb, Anti-Jsb, and other high-incidence Kell
DISEASE ASSOCIATION
Antigens
Anti-I & Pathogenic autoanti-I
- Rarely seen due to the few individuals who lack
- Production is stimulated by microorganisms with I-
these antigens
like antigen on their surface like Mycoplasma
pneumoniae, and Listeria monocytogenes
Anti-i KX BLOOD GROUP SYSTEM (019)
- Infection with Epstein-Barr virus, reticuloses, - Present on all RBCs except those of rare McLeod
myeloid leukemias, and alcoholic cirrhosis phenotype
- IgG anti-i has also been described and has been Kx Antigen
associated with HDN - Increases when Kell antigens are denatured
- Might be a precursor or backbone for Kell
KELL (006); KEL; C-7 &
KX BLOOD GROUP SYSTEM (019); XK; C-Xp DISEASE ASSOCIATION
→ The first group detected after the introduction of Anti-K
antiglobulin testing - Naturally occurring Anti-K IgM class is rare and have
→ First detected in the serum of Mrs. Kellaher been associated with bacterial infections
→ The antibody detected reacts with the RBCs of her - A 20 day-old infant with an Escherichia coli
newborn infant, her older daughter, her husband, 0125:B15 infection whose mother did not make any
and about 7% of random population anti-K
→ Anti-k (Cellano)- high incidence antithetical partner - Other organisms include 1Mycobacteria,
2
to K Enterococcus faecalis, 3Morganella morganii,
4
Campylobacter jejuni, and 5Campylobacter coli
DUFFY BLOOD GROUP (008); ACKR1; C-1
ANTIGENS
K and k antigen - Named after Mr. Duffy, a multi-transfused
- K is rated as second in immunogenecity and a rare hemophiliac which anti-Fya was found
antigen - In serum of woman who had three pregnancies
- k is more common than K found the antithetical partner which was Fyb
Kpa, Kpb, and Kpc antigens
- Kpa and Kpc are low-incidence partner of Kpb ANTIGENS
- Kpa gene is associated with suppression of other Kell Fya and Fyb Antigens
antigens on the same molecule, including k and Js - Two most important Duffy antigens in routine
- Kpc antigen found in siblings from a consanguinaeous blood bank
married couple in Japan who type Kp(a-b-) but have - They can be identified on fetal RBCs as early as 6
normal Kell antigens weeks gestational age and are well developed at
Jsa and Jsb Antigens birth
- Jsa antigens is antithetical to the high-incidence - These antigens have not been found on platelets,
antigen Jsb and WBCs
- Linked to Kell system after the discovery of Ko RBCs - Can be found in brain, colon, endothelium, lung,
were Js(a-b-) spleen, thyroid, thymus and kidney cells
Other Kell Antigens - Do not store well on saline suspension and tend to
- K10, K11, K12, K13, K14, K18, K19, K22, K23, K24, elute from RBCs stored in a medium with low pH or
K25, K26, AND K27 low ionic strength
McLeod Phenotype - Destroyed by common proteolytic enzymes:
- Occurs in males and inherited from a carrier mother o Ficin
o Papain
ANTIBODIES o Bromelain
Anti-K o Chymotrypsin
- Most common antibody seen in the blood bank
aside from ABO and Rh antibodies ANTIBODIES
Malaika Ina M. Galos BSMLS-SP1 18
Anti-Fya and Anti-Fyb Lua and Lub
- Anti-Fya is a common antibody and is found as a - Antigens produced by allelic codominant genes
single specificity or in a mixture of antibodies - Antigens have been detected on fetal RBCs as early
- Anti-Fya occurs three times less frequently than anti- as 10-12 weeks of gestation
K - They are poorly developed at birth
- Anti-Fyb is 20 times less common than anti-Fya and - Do not reach adult levels until the age of 15
often occurs in combination with other antibodies - Not detected on platelets and WBCs
- These antibodies are usually IgG and react best at - Lutheran glycoprotein is widely distributed in
antiglobulin test tissues of the 1brain, 2pancreas, 3placenta, 4skeletal
- Enhanced in low ionic strength medium muscles, 5hepatocytes (especially fetal hepatocytes)
- Some of the antibodies show dosage
ANTIBODIES
DISEASE ASSOCIATION Anti- Lua
- Fy(a-b-) is associated with resistance to in vitro - Mostly are IgM in class
infection by Plasmodium knowlesi and Plasmodium - Unusual class of the Lutheran antibodies is the IgA
vivax class
- Not profoundly altered with the common blood
KIDD BLOOD GROUP (009); SLC14A1; C-18 bank enzymes ficin and papin, but it can be
destroyed by trypsin, chymotrypsin and pronase
→ Simplest and most straightforward system Anti-Lub
→ Found primarily in the serum of Mrs. Kidd, a mother - Mostly are of IgG class
whose infant had erythroblastosis fetalis - Some are IgM and IgA
→ It has special significance in routine blood banking - Alloanti- Lub reacts with all cells tested except the
due to difficulty in the detection of its antibodies autocontrol, and reactions are often weaker with
and is one of the common cause of HTR Lu(a+b+) RBCs and cord RBCs
- Ficin and papain does not significantly alter
ANTIGENS reactivity
Jka and Jkb
- Common RBC antigens ✓ A major blood group system outside ABO and Rh
- Well developed in RBCs of the neonates become important only after patients develop
- Jka has been detected on fetal RBCs as early as 11 unexpected antibodies
weeks ✓ Fundamental knowledge of antibody characteristics,
- Jkb has been detected at 7 weeks, this early clinical significance, and antigen frequency are
development of antigens contributes to the needed to help confirm antibody specificity and to
potential for HDN select appropriate units for transfusion
- Not very immunogenic
- Not denatured by papain or ficin
- Antigens are not found on platelets, and WBCs

ANTIBODIES
Anti- Jka and Anti- Jkb
- Have notorious reputation in blood banking
- Demonstrates dosage, often weak, and are found in
combination with other antibodies which make them
difficult to detect
- Anti- Jka is more common than anti- Jkb and they are
usually of IgG class

LUTHERAN BLOOD GROUP (005); BCAM; C-19

→ Anti- Lua is discovered in patients with lupus
erythematosus diffusus following the transfusion of
a unit of blood carrying the corresponding low-
incidence antigen
→ Named after the name of the donor Luteran

ANTIGENS
Malaika Ina M. Galos BSMLS-SP1 19
WEEK 6 (Lecturer: Sir Edgardo Monisit) 3. Add 1-2 drops of antihuman globulin reagent
4. Centrifuge for 20 seconds at 1000 RCF
5. After centrifugation, completely resuspend the cell
ANTIGLOBULIN TEST
pellet by gently tipping and rolling the tube
Additional knowledge 6. Read for agglutination macroscopically with the aid
• Anti-A Reagent- Trypan Blue/ Patent Blue of a background light source
• Anti-B Reagent- Acriflavine/ Tartazine 7. Incubate the tube to 5 minutes at room
temperature
Antiglobulin Testing 8. Centrifuge the tube for 20 seconds at 1000 RCF
- Discovered in 1945 9. Completely resuspend the cell pellet by gently
- Otherwise known as “coomb’s test” tipping and rolling the tube and read for
- Based on the principle that antihuman globulins agglutination.
(AHGs) obtained from immunized nonhuman
species bind to human globulins
INDIRECT ANTIGLOBULIN TESTING
▪ Performed to determine in-vitro sensitization of
TYPES OF AHG REAGENTS
RBCs
• Polyspecific AHG- target both IgG and complement ▪ Used in:
• Monospecific AHG- target either IgG or complement o Incomplete antibodies to potential donor
RBCs or to screening cells in serum
Antihuman Globulin Reagent Components o Determination of RBC phenotype using
• Anti-IgG (IgG1, IgG2, IgG3, IgG4) known antisera (ex. Weak D)
• Anti-complement (C3b, C3d, C4b, C4d) o Titration of incomplete antibodes
▪ Add check cells to check if the reagent is working
REAGENT PREPARATION
Procedure
1. Combine sera and cells. Either the sera or the cells
Polyclonal AHG production- recognize different/multiple Ag
comprised the known factor in the test scheme
epitope
2. Centrifuge tubes
- Usually prepared in rabbits, although when large
3. Examine, interpret and record
vol. Of antibody are required, sheep or goats may be
4. Add enhancement media
used
5. Incubate for the appropriate time designated by the
- Reacts across all complemet fragments of IgG
enhancement media, do not incubate beyond the
component
enhancement media as the Ag-Ab- complexes may
- Anti-IgG (IgG1, IgG2, IgG3, IgG4)
begin to dissociate
- Anti-complement (C3b, C3d, C4b, C4d)
6. Centrifuuge
7. Examine, interpret, record
Monoclonal AHG production- recognize/reacts to a single Ag
8. Wash tubes 3 times
epitope
9. Add AHG sera
- Presented by Kohler and Milstein
10. Centrifuge the tubes
- Used to produce AHG and has proved particularly
11. Examine, interpret, record the result. If negativ,
useful in producing high-titer antibodies with well-
incubate for 5 minutes and re-examine
defined specifities to IgG and to fragment of C3
12. Add check cells to all negative tubes
13. Centrifuge
TYPES OF ANTIGLOBULIN TESTING 14. Examine, interpret, record

DIRECT ANTIGLOBULIN TESTING FACTORS AFFECTING ANTIGLOBULIN TESTING

▪Detects in-vivo sensitization of RBCs w/ IgG and/or → Ratio of serum to cells
complement → Reaction medium
▪ Conditions that can result in in vivo coating of RBC → Temperature
o Hemolytic Disease of the Newborn → Incubation time
o Hemolytic Transfusion Reaction → Washing of RBCs
o Autoimmune Hemolytic Anemia → Saline for washing
Procedure (NOTE: does not require incubation phase because of the Ag-Ab complexes → Addition of AHG
formed in vivo) → Centrifugation for reading
1. From washed RBC, make a red blood cell suspension
2. Add 1 drop of RBC from the RBC suspension and
SOURCES OF ERROR
wash the cells 3 times

Malaika Ina M. Galos BSMLS-SP1 20

FALSE-POSITIVE RESULTS • Screening for unexpected antibodies to red cell
▪ Improper specimen antigens
▪ Autoagglutinatable cells • Antibody identification if unexpected antibodies
▪ Dirty glassware are detected
▪ Bacterial contamination of cells or saline used in • Comparison of current and previous test results,
washing and any discrepancies shall be investigated, and
▪ Cells with a positive DAT used for the IAT appropriate action taken before a unit is issued
▪ Saline contaminated by heavy metals or colloidal for transfusion
silica 4. Donor RBC unit testing:
▪ Overcentrifugation or overreading • ABO group confirmation and Rh type
▪ Polyagglutinable cells confirmation for Rh-negative RBC units
▪ Preservative dependent antibody in LISS reagents 5. Donor red blood cell unit selection:
(IAT) • Selection of components of ABO group and Rh
▪ Contaminating antibodies in the AHG reagent type that are compatible with the transfusion
▪ Centrifugation of test with polyethylene glycol prior recipient and with any unexpected allogeneic
to washing antibodies
6. Compatibility testing (crossmatch):
• Serologic
FALSE-NEGATIVE RESULTS • Computer or electronic
▪ Inadequate or improper washing of cells
7. Labeling of blood or blood components with the
▪ AHG reagent nonreactive because of deterioration
recipient’s identifying information and issue
or neutralization
▪ AHG reagent not added
▪ Serum not added in the IAT
▪ Cell suspension either too weak or too heavy CROSSMATCHING
▪ Low pH of saline
▪ Poor reading technique
SEROLOGIC CROSSMATCH
▪ Undercentrifuged or overcentrifuged
▪ Consists of mixing the recipient’s sample/ donor’s
sample
MODIFIED & AUTOMATED ANTIGLOBULIN ▪ Several procedures can be used including:
→ Low Ionic Strength Polybrene Technique o Immediate-spin crossmatch
→ Enzyme-Linked Antiglobulin Test o Antiglobulin crossmatch
→ Solid-phase Technology
→ Gel Test A. Immediate spin crossmatch
o Neutral- acts as a trap - Mixing of donor RBCs w/ the recipient’s serum and
o Specific- uses specific IgG incorporated into spun
the gel - Does not detect all ABO incompatibilities
o Antiglobulin- AHG reagent is incorporated
in the gel • Type and screen- blood type then Ab screen
• Abbreviated crossmatch- type and screen w/ an
immediate spin crossmatch
COMPATIBILITY /PRETRANSFUSION TESTING
B. Antiglobulin crossmatch
• Serologic test done before transfusion to confirm - Begins in the same manner as immediate spin
that blood unit is sustainable for transfusion o Add the sample,centrifuge,continue to the
• All entire quality process composed of many 37C phase, then AHG phase
procedures designed to provide the safest blood
product possible for the recipient of a transfusion Interpretation
- Tubes are examined for hemolysis
Steps in Pretransfusion Testing - Presence of hemolysis/ agglutinable inidcates
1. Request for transfusion incompatibility
2. Identification of transfusion recipient and blood - Absence of hemolysis/ agglutination inidcates
specimen collected compatibility
3. Testing of transfusion recipient’s blood specimen:
• Evaluation of specimen for testing suitability
• ABO group COMPUTER CROSSMATCH
• Rh type (Weak D testing is optional when ▪ Indicate that an electronic crossmatch to detect
testing the patient) ABO incompatibilities

Malaika Ina M. Galos BSMLS-SP1 21

CAUSES OF POSITIVE RESULTS IN THE
SEROLOGIC CROSSSMATCHING
▪ Incorrect ABO grouping
▪ Abnormalities in the patient’s serum
▪ An alloantibody
▪ Autoantibodies
▪ Prior sensitization (pregnancy)
▪ Contamination in test system
▪ Selection of appropriate ABO/Rh donor compatible
to the recipient
▪ Prior coating of the donor RBCs w/ protein

Other Transfusion Performed

• Transfusion of plasma products- crossmatching is
not required (NOTE: minor crossmatch if more than
10 units)
• Intrauterine transfusion- crossmatching is required
o Blood unit must be the same with mother’s
blood type
• Neonatal transfusion- crossmatching is required
o Maternal blood sample is used
• Massive transfusion- crossmatching is required
(would yield false results)
• Autologous transfusion- crossmatching may be
required
• Platelets- crossmatching is not required

Malaika Ina M. Galos BSMLS-SP1 22

WEEK 7 (Lecturer: Edgardo Monisit)
ADVANTAGES DISADVANTAGES
- Decreased disease transmission - Bacterial Contamination
DONOR SELECTION - Decreased transfusion reaction - Circulatory Overload
- Decreased alloimmunization - Cytokine-mediated reaction
• Encompasses the medical history requirements of - Blood product misidentification
the donor, the partial physical examination, and
serologic testing of the donor blood

Information Required for the Registration of Donor

METHODS USED TO OBTAIN AUTOLOGOUS
1. Name BLOOD
2. Date
3. Address Preoperative Collection
4. Telephone number - 5-6 weeks before scheduled operation
5. Gender - Last blood collection should be no later than 3 days
6. Age and date of birth - Usually performed in surgeries like: Orthopedic,
7. Consent to donate Vascular, Cardiac, Radical prostatectomy
- Women before pregnancy
Medical History Questionnaire
- Standardized questionnaire by AABB, FDA, blood & Acute Normovolemic Hemodilution
plasma industry - Maintains blood volume
- Self-administered questionnaires must be reviewed - Crystalloids 3:1, Colloids 1:1
by trained personnel before completing the - Recommended that a px start with <12 g/dL
screening process and prior to collecting blood - Perfromed in surgery & managed by
- Answerable by “yes” or “no” ANESTHESIOLOGY
- Must be answered by the day of the donation - Transfused w/in 8 hours
- Reinfused in reverse order from collection
PHYSICAL EXAMINATION
Intraoperative Blood Collection
1. General appearance - Involves collecting shed blood from the surgical site
2. Weight - The procedure is contraindicated if there is potential
3. Temperature fro contamination of the surgical site (ex.
- Ask not to drink coffee or hot beverage Appendectomy)
- Oral temperature are lower than normal (not
cause for deferral) Postoperative Salvage
4. Pulse - Collected from a drainage tube placed at the surgical
- Should be counted for 15 seconds (multiply by site
4) - Blood must be reinfused w/in 6 hours
5. Blood Pressure - Should be no more than 1.4 L
- 140 / 100 (acceptable blood pressure in pH)
6. Hemoglobin
- Allogenic: 12.5 g/dL DIRECTED DONATION
- Autologous: 11 g/dL → unit collected is directed toward a specific patient
7. Hematocrit → unit must be irradiated to prevent TA-GVHD
- Allogenic: 38%
- Autologous: 33% APHERESIS
→ collecting a specific blood component while
SEROLOGIC TESTING returning the remaining whole blood components
back to the patient
ABO/ Rh Typing Screening Tests → most commonly guaranteed platelet collection
Antibody Screening Hepatitis B/C
Malaria
Syphilis
HIV
PLATELETPHERESIS
HbsAg ▪ otherwise known as “single donor platelet”
Anti-HBc ▪ >75% of platelet transfusion are pheresis-derived
Anti-HCV ▪ 1 unit= 6-8 random donor platelet
Anti-HIV ½
Anti-HTLV
▪ Single product- therapeutic dose for adult platelets
▪ Exposed to a single set of platelet antigens
▪ Interval is at least 2 days, not to exceed more than
AUTOLOGOUS DONATION twice a week or more than 24 times a year
Malaika Ina M. Galos BSMLS-SP1 23
PLASMAPHERESIS → A positive test result at any phase of testing
▪ 1st product to be collected by apheresis inidcates the need for antibody identification
▪ Used to collect transfusable fresh-frozen plasma studies
▪ Donors are classified as “infrequent” or “serial” QUESTIONS TO CONSIDER:
1. In what phase(s) did the reaction(s) occur?
- IS phase- IgM
LEUKAPHERESIS - 37C incubation phase- warm reacting
▪ Only effective method for collecting leukocytes, antibodies
more specifically, granulocytes - AHG phase- IgG/ non-agglutinating
▪ It is only indicated for those people who has been antibodies
infected with antibiotic/ antimicrobial resistant 2. Is the autologous control negative or positive?
strains of microorganisms - Negative- proceed to the testing
▪ Prevents: 1TA-GVHD, 2T cell proliferation, 3Febrile - Positive- discard (the px may have
Non-Hemolytic Transfusion Reaction autoantibodies)
3. Did more than one screen cell sample react? If so,
DOUBLE RBC PHERESIS did they react at the same strength and phase(s)?
▪ Double units of red blood cells collected using - 1+ (small tiny agglutinates)
apheresis equipment - 2+ (medium sized agglutinates)
▪ Used to collect allogenic & autologous units - 3+ (several large agglutinates)
- 4+ (one solid agglutinate)
4. Is hemolysis or mixed field agglutination present?
ANTIBODY SCREENING - Hemolysis-there is complement activation
• Ambot paspas kays sir basa nalang tas libro - Mixed field- the Abs are non-complement
• Screen cells activating
o antigens which can possibly be present in 5. Are the cells truly agglutinated, or is roleaux
the donor’s red blood cells present?
o should be well represented of each of the
cell. It could be 3-5 screen cells ANTIBODY SCREENING
i. Observe the results in the AHG phase
TECHNIQUES OF THE ANTIBODY SCREEN ii. Perform check cell to those that are negative
iii. Compare the results of the AHG to the panel cells
whether it is positive (+) or negative (-)
TUBE TECHNIQUE a. If (+) ang AHG automatic cancel the (-) on
▪ Conventional technique the panel cells
▪ Technique where DAT is performed b. If (-) ang IS, 37, AHG cancel all the (+) on
▪ Patient’s serum or plasma is tested with Group O the panel cells
RBCs with known antigens Rules
▪ To observe for agglutination/ hemolysis in the IS -in the Ag side (panel cells), (+) sign indicates the presence of the Ag, (-)
phase, 37C phase, AHG phase sign denotes absence of Ag
▪ Hemolysis= presence of antibody against the
antigen on the red blood cell

GEL TECHNIQUE
▪ Uses gel as a trap
▪ Eliminate washing
▪ LISS as diluent (.8%)
▪ Contains IgG

SOLID-PHASE
▪ RBC antigens coat microtiter wells instead of being
present on the packed RBCs
▪ Patient serum is added to each well on the screen
cell set with LISS
▪ Anti-IgG (on the cells)

INTERPRETATION OF RESULTS

Malaika Ina M. Galos BSMLS-SP1 24

WEEK 8 (Lecturer: Doc Eleanor Apita) eligible to donate by apheresis after the
normal deferral period (2 days-plasma, 8
wks.-hemapheresis)
TRANSFUSION MEDICINE
• Is a multidisciplinary specialty encompassing all Therapeutic hemapheresis
aspects of blood donation, blood component - is the removal of harmful cellular or plasma factors.
preparation, blood cell serology and blood
transfusion therapy

Operationally, Transfusion medicine is divided between:

• Blood centers
o Recruits and collect blood from donors and
manufacture and distribute blood
components
• Transfusion services
o Perform pre-transfusion compatibility
testing, select and issue blood components TYPES OF TRANSFUSION
for patients and provide medical support
for blood transfusion
AUTOLOGOUS
▪ A recipient receives his/her own blood
Blood transfusion
▪ Prevention of transfusion-transmitted diseases
- is the administration of whole blood or a component
▪ Elimination of alloimmunization
to replace blood lost through trauma, surgery or
▪ Elimination of GVHD
disease
▪ Erythropoietic stimulation
▪ Safest blood a recipient can receive
Apheresis
- removing whole blood from an individual,
manipulating the removed blood, and subsequently INTRAOPERATIVE SALVAGE
re-infusing portions of the blood ▪ A system where blood is aspirated from the
operative field and then transfused to the patient
Hemapheresis ▪ Transfusion is done directly using a microaggregate
- is a procedure in which WB is removed from an filter or washed to remove debris, phospholipids,
individual, anticoagulated, and separated into activated complement and tissue activators
components ▪ 70-260 um pore size
- process of removing normal or abnormal
constituents (polycythemia vera) from circulating
blood DIRECTED TRANSFUSION
- therapeutic phlebotomy ▪ Patient directly solicits blood from family and friends
▪ The process is based on false assumption that blood
Cytapheresis donated by family and friends is safer than that of a
- is the removal of the cellular components of blood regular volunteer donor
- RBC, platelets, leukocytes
MASSIVE BLOOD TRANSFUSION
Plasmapheresis ▪ When a volume equivalent to patient’s blood
- removal of plasma and all plasma constituents volume is transfused within a 24-hour period.
o donor requirements is the same as WB ▪ Transfusion of 16-20 units (10 units Harmening) of
donation RBCs
o donors can donate as often as twice per
week but cannot donate more than 24
times per year (hemapheresis & NEONATAL TRANSFUSION
plateletpheresis) ▪ To replace blood drawn for laboratory tests and to
o individuals donating after whole blood treat anemia of prematurity.
donation or after an instrument failure ▪ Small aliquots of donor blood can be transferred
resulting in the inability to return the from the collection bag to a satellite bag or transfer
extracorporeal blood volume , must wait 8 bag
weeks before donating by apheresis ▪ Blood can be withdrawn from the collection bag or
o if the extracorporeal blood volume of the transfer bag using an
instrument is less than 100 mL or if the loss o injection site coupler and needle and
of the RBC is less than 200 ml the donor is syringe or
Malaika Ina M. Galos BSMLS-SP1 25
 o sterile docking device and syringe. - Compatibility with the patient’s recorded blood type
▪ Unit must be less than 7 days old (14-21 days old at should be verified
some institutions) to reduce hyperkalemia and to - The expiration time and date of the blood
maximize 2,3-DPG. component must be verified as acceptable
- Recheck the unit/ component to be transfused
- The physician’s order and patient’s consent for
EXCHANGE TRANSFUSION
transfusion should also be verified
▪ Removes unbound IgG antibodies, bilirubin and
- 70-260 um pore size filter (reduce CMV)
antibody-coated RBCs.
- Leukoreduction filter maybe used (RBC & PC)
▪ Corrects anemia and replaces fetal RBCs with adult
RBCs w/c have better release of oxygen to the
Administration sets should contain the ff:
tissues
- Drip chamber
- Attached compatible intravenous solution
EMERGENCY TRANSFUSION - Means of controlling the flow rate
▪ Patients who are rapidly or uncontrollably bleeding - Microaggregate filter is used in surgery
may require immediate transfusion. - Normal saline is preferred solution for all
▪ Group O RBCs (packed cells) are selected transfusions
▪ Group O negative if patient is a female of - Calcium-containing solutions should be avoided as
childbearing age these may precipitate clotting
▪ An Rh negative male can be switched to Rh positive - RBC should not be administered with 5% dextrose to
if few O negative units are available or if massive avoid hemolysis
transfusion is required. - Medications should not be added to blood
components

TRANSFUSION IN ONCOLOGY
▪ Irradiated blood components to avoid TA-GVHD BLOOD COMPONENTS AND DERIVATIVES
(Transfusion Associated Graft versus Host Disease)
▪ For cancer patients
WHOLE BLOOD
▪ Massive transfusion where both red cell mass and
plasma volume are required
Speed of infusion ▪ Replacement of the O2 carrying capacity and
- 15 drops = 1ml maintenance of the intravascular volume
- At a rate of 60 drops/min, 60/15 x 60 = 240 ml of ▪ Used in active brisk bleeding
blood can be transfused in 1hr ▪ A product in w/c all of the red cells & most of the
- 1 unit can be transfused in 2 hours plasma from the original unit remain
- Slow infusion w/in 4 hours ▪ 540 ml
- Platelet or plasma 30-60 minutes ▪ Shelf life: CPD – 21 days; CPDA1 – 35 days at 1-6°C

Blood warmers
RED BLOOD CELLS
- Must be used to avoid arrhythmia or death of the
▪ Packed RBC with reduced plasma (avoid circulatory
patient
overload) volume
▪ May be prepared from whole blood by overnight
Transfusion Administration
sedimentation or by centrifugation using heavy spin
- Physician’s order is required to prepare, dispense,
▪ To increase O2 carrying capacity of blood in
and administer blood components
patients with acute/ chronic anemia
- 18 gauge needle or catheter is sufficient
▪ Who are symptomatic (syncope, dyspnea) or at high
risk for end organ damage (subcritical coronary
Prior to Transfusion, the following must be done:
atherosclerosis)
- Identification of the component
▪ 260 ml
- Identification of the patient
▪ Shelf life: same as whole blood
- At least two unique identifiers
▪ Name
▪ Registration number RED BLOOD CELLS IN ADDITIVE SOLUTION
- The unit identifier on the blood container should be (ADENINE-SALINE)
checked against the associated documentation ▪ RBCs with reduced plasma volume and an additional
*(transfusion form or attached tag) of 100ml additive solution
- ABO and Rh type on the unit must agree with the ▪ Same as whole blood or packed RBC
associated documentation ▪ 340 ml

Malaika Ina M. Galos BSMLS-SP1 26

 ▪ Shelf life: 42 days at 1-6°C ▪ 200-400 ml
▪ Shelf life: 5 days at 20-24°C with constant gentle
WASHED RBCS ( NO PLASMA ) agitation
▪ Prevention of recurrent or severe allergic reactions
▪ Neonatal intrauterine transfusion LEUKOCYTE REDUCED PLATELETS
▪ Prevention of anaphylactic reactions in IgA-deficient ▪ Platelets, random donor or pheresis
patients ▪ Indications, as per random donor or pheresis
▪ 180ml platelets
▪ Shelf life: 24 hours at 1-6°C ▪ When additional prevention of alloimmunization to
HLA antigens or reduction of CMV infection risk is
FROZEN DEGLYCEROLIZED RBCS desired
▪ Removes >90% of WBC ▪ Vol. similar to original component
▪ No plasma ▪ Shelf life: similar to original component when closed
▪ Long-term preservation of RBC units with rare system is used
phenotype or autologous RBC units ▪ Bedside filtration is done immediately before
▪ Alternative to CMV-seronegative units in transfusion (3log filters)
emergencies ▪ 3rd gen filter
▪ Decreases febrile transfusion reactions
▪ 250 ml FRESH FROZEN PLASMA
▪ Shelf life: 10 years at -65°C to -200°C & 24 hours at ▪ When there is a specific coagulation factor
1-6°C after washing (AG:40% glycerol) deficiency where coagulation factors are not
available
LEUKOCYTE-REDUCED RBCS ▪ All coagulation factors, complement, other plasma
▪ Decreases risk of alloimmunization to HLA antigens proteins
on leukocytes ▪ Bleeding due to multiple coagulation factor
▪ Decrease febrile non-hemolytic transfusion reactions deficiency (liver failure, DIC, coumadin toxicity)
▪ Reduce the risk of CMV infection ▪ Vol 200-260 ml
▪ Decrease immunomodulation, and may decrease ▪ Shelf life: 1 year at -18°C or below
the risk of prion transmission in B lymphocytes
▪ 3 log filtration CRYOPOOR PLASMA
▪ Vol. similar to original unit ▪ Component remaining after cryoprecipitate is
▪ Shelf life: similar to original unit when closed system prepared
used ▪ Minimal concentrations of fibrinogen, factor VIII
and vWF
TWO-UNIT RBCS, APHERESIS ▪ Used in patients with hemolytic uremic syndrome,
▪ Two RBC is divided into two separate RBC units of thrombotic thrombocytopenic purpura
equal volume ▪ Vol. 200ml
▪ Similar to pRBC (increase oxygen carrying capacity) ▪ Shelf-life: same as FFP 1 year at -18°C or below
▪ Same indications as pRBC
▪ Vol. 320 ml CRYOPRECIPITATE
▪ shelf life: 1 year at 1-6 °C ▪ Derived from FFP
▪ For acquired deficiencies of fibrinogen and factor
RANDOM DONOR PLATELET CONCENTRATE XIII deficiency
▪ Bleeding due to quantitative and qualitative platelet ▪ Used in vWF and hemophilia A when virus-
disorders inactivated factor concentrates are not available
▪ Prophylaxis in severely thrombocytopenic patients ▪ Maybe beneficial in bleeding due to uremic platelet
(5,000 – 10,000/uL) dysfunction not responding to desmopressin,
▪ Vol. 50-70 ml estrogens
▪ Shelf life: 5 days at 20-24 °C with constant gentle ▪ Factor XIII deficiency, unresponsive to dialysis
agitation & 4 hours after pooling ▪ Vol. 10-15 ml
▪ Shelf-life: 1 year at -18°C or below and 6 hours after
thawing
PLATELETS, PHERESIS (SINGLE DONOR
PLATELETS)
▪ Platelets in approx. 250ml of plasma to maintain pH
FACTOR VIII (LYOPHILIZED)
▪ Available in several degrees of purity
above 6.2
▪ All products are lyophilized
▪ Indications, similar to random donor platelets
▪ Fractionation of pooled donor plasma
Malaika Ina M. Galos BSMLS-SP1 27
 ▪ Moderate to severe Hemophilia A ▪ indicate adverse results when used in critically ill,
▪ Vol. 25 ml of sterile diluent for reconstitution trauma, or elderly patients
▪ Shelf-life: 2 years at 2-8°C ▪ contraindicated in dehydrated patients
▪ flushing, urticaria, chills, fever, and headache may
FACTOR IX occur
▪ Plasma-derived and recombinant proteins are ▪ 25-50 ml
available ▪ Shelf life: 5 years at 2-10°C
▪ Factor IX complex: factors II, VII, IX and other
proteins PLASMA PROTEIN FRACTION
▪ Factor IX, human purified by immunoaffinity ▪ Contains 83% albumin and 17% globulin
chromatography from pooled human plasma ▪ Indications, similar to albumin
contains other traces of proteins ▪ Hypotension may occur when given at rates of over
▪ Recombinant factor IX 10 ml/min
▪ Vol of 25 ml of sterile diluent for reconstitution ▪ 25-50ml
▪ Shelf life: 2 years at 2-8°C ▪ Shelf life: 3 years below 30°C
▪ Plasma derived factor IX complex is used to treat
hemophilia B and in patients with inhibitors to factor IMMUNE GLOBULIN, INTRAVENOUS
VIII or IX ▪ Mostly IgG antibodies, traces of IgA and IgM
▪ Prepared from pooled human plasma by cold
ANTITHROMBIN III CONCENTRATE ethanol fractionation
▪ Prepared from pooled human plasma by modified ▪ Primary humoral immunodeficiency, ITP, bone
cold ethanol fractionation marrow transplant and pediatric HIV infection
▪ Treatment or prophylaxis of thrombotic events in ▪ It has gained acceptance in the treatment of
patients with congenital antithrombin deficiency Guillain-Barre’s syndrome
(DIC, heparin resistance, sepsis, deep vein ▪ 10-20ml
thrombosis, pulmonary embolism) ▪ Shelf life: 3 years at 2-8°C
▪ Vol. per manufacturer’s directions
▪ Shelf life: 2 years at 2-8°C RH IMMUNE GLOBULIN
▪ IgG anti-Rho, full dose is 300ug, minidose is 50ug
S/D (SOLVENT/DETERGENT) TREATED PLASMA ▪ Prevention of RH alloimmunization of RH negative
▪ Plasma similar to contents as in FFP individual exposed to the D antigen
▪ May have minimal content of high-molecular ▪ Used to prevent Hemolytic Disease of the Newborn
weight vWF ▪ 1ml
▪ S/D treatment of plasma is to inactivate lipid- ▪ Shelf life: 1 ½ years at 2-8°C
enveloped viruses (HIV, HCV)
▪ Indication, similar to FFP FIBRIN SEALANT
▪ 200ml ▪ Fibrinogen, human thrombin, aprotinin and calcium
▪ Shelf life: 1 year at -18°C from manufacturing chloride
▪ Topical application under direct visualization
DONOR-RETESTED FROZEN PLASMA ▪ Adjunct to hemostasis in cardiopulmonary bypass
▪ Plasma, as in FFP, stored and released for use after surgery
donor has been retested negative for all viral ▪ Sealing anastomoses, treatment of spleen injuries
markers after 112 days or more, allowing for ▪ Adjunct to surgical methods of hemostasis
window period infections to seroconvert
▪ Indications, similar to FFP Centrifugation speed
▪ Lower risk of viral transmission than FFP
▪ 200-260 ml - 5000 rcf for 5 mintues PRBC/platelet concentrate
▪ Shelf life: 1 year at -18°C then centrifugation is @ room temperature (20-24C)
- 5000 rcf for 7 minutes cryoprecipitate of cell free
ALBUMIN plasma (heavy spin)
▪ 96% albumin, 4% other plasma proteins - 2000 rcf for 3 minutes platelet-rich plasma (light
▪ Prepared by cold ethanol fractionation spin)
▪ Indications are controversial, as it increases colloid-
oncotic pressure, maintaining blood volume
transiently
▪ hemolysis has been reported to occur when it is
diluted in water and infused in the same line as RBCs
Malaika Ina M. Galos BSMLS-SP1 28
WEEK 9 (Lecturer: Doc Eleanor Apita) → reaction will take 1-2 hours. signs & symptoms can
occur w/in mins. after starting the transfusion
TRANSFUSION REACTIONS → common Abs that cause IHTR: Anti-A, Anti-K, Anti-
Jka, Anti-Fya
• Diverse group of adverse reactions to transfusion → Immune-Mediated IHTR can destroy RBCs by one of
that usually present during or shortly after two mechanisms: Intravascular & Extravascular
transfusion Hemolysis
• If a transfusion reaction occurs while a transfusion is
in progress, the transfusion should be stopped INTRAVASCULAR HEMOLYSIS - Intravascular RBCs lysis
immediately, and the intravenous line should be releases 1Hgb, 2RBC stromata and 3cellular enzymes
kept open with saline manifesting hemoglobinemia & hemoglobinuria
• Suspected transfusion reactions are underreported Affected organs:
in passive surveillance systems • Kidney- damage to sclera of glomerulus, cortex &
• Transfusion is an irreversible event that carries tubules
potential benefits and risks to the recipient • Liver- damage to hepatic portals and hepatocytes
• Any unfavorable transfusion-related event occurring EXTRAVASCULAR HEMOLYSIS- characterized by Ag-Ab
in a px during or after transfusion of blood complexes formation of RBCs incomplete activation of
component complement
Signs & Symptoms- usually mild and not life-threatening
Rates of Risk • fever
1. NHFTR 1-2% • chills
2. Allergic Transfusion Reactions 1-2%
• jaundice
3. IHTR & DHTR 1:6,000
• unexpected anemia
4. Fatal immediate 1:100,000
• decreased haptoglobin
Therapy & Prevention
Occurrence of Reactions and Fatality Episodes
- extravascular IHTR does not require therapeutic
• AHTR 1:25,000
intervention, vital signs, coagulation status and renal
• DHTR 1:2,500
output
• Noncardiogenic Pulmonary Edema/TRALI 1:1,000 - all policies & procedures should be followed to
• TA-GVHD & related donor 1:7,000 ensure proper patient identification, handling &
• TA-GVHD & unrelated donor 1:39,000 correct transfusion at the bedside

Errors Associated with Transfusion Reactions

- Improper specimen identification
- Improper patient identification Signs & Symptoms of IHTR: (observed in a conscious px)
- Antibody identification error • fever w/o chills
- Crossmatch procedure error • coagulopathy
• oliguria
HEMOLYTIC TRANSFUSION REACTION • anuria
• death w/ sustained hypotension
→ can occur either at the time of transfusion
(immediate) or a few days 3-7 days after transfusion Therapy & Prevention
(delayed) - monitor the px for DIC, hypotension & acute renal
→ HTR oftern occurs: failure
- Transfusion of ABO-incompatible RBCs - mannitol is used to induce renal diuresis and to
- Transfusion of plasma containing products prevent renal failure
(plasma&platelets) (leukocytes from the - IV fluids is used to treat hypotension & vasoactive
donor’s blood) drugs
- Physically or chemically induced - component therapy for px w/ bleeding diathesis of
coagulation abnormalities
IMMEDIATE HEMOLYTIC TRANSFUSION
REACTION DELAYED HEMOLYTIC TRANSFUSION REACTION
→ occurs very soon after the transfusion of → most often the result of an anamnestic response in a
incompatible RBCs px who has previously been sensitized by transfusion
→ RBCs are rapidly destroyed releasing Hb& RBC reaction, pregnancy, or transplant
stromata into the circulation → clinical signs and symptoms are usually mild
→ hemoglobinuria & hemoglobinemia → severe DHTR cases and fatalities are uncommon

Malaika Ina M. Galos BSMLS-SP1 29

 → unexpected or unexplained decreases in Hgb or Hct - bedside filtration, washed RBCs, deglycerolized is
values post transfusion should be investigated as a indicated
possible DHTR - antipyretics (fever)
→ maybe associated w/ bone marrow transplantation
→ TYPES: ALLERGIC TRANSFUSION REACTION
o Secondary- as an anamnestic response to
→ commonly reported as FNHTR
transfused RBCs. 3-7 days from the
→ appear w/in minutes during blood transfusion
transfusion to cause extravascular
(immediate hypersensitivity type)
hemolysis
→ histamine is the primary mediator
o Primary alloimunization- the first time the
px has been transfused → Etiology
o Donor plasma has allergens that has IgE or
Signs & Symptoms and Clinical Work-Up IgG that reacts with the patient’s antigen
o The donor plasma has reagins IgE or IgG
• mild, complement is not activated (no intravascular
both that combine with allergens in px
hemolysis)
plasma
• mild fever (common manifestation)/ chills
• moderate jaundice
Signs & Symptoms
• oliguria and DIC are rare
• erythema
• blood specimen should be sent to the blood bank
• hives
for PTT reaction investigation
• pruritus
• tests for hb, hct coagulation studies and renal
function should be done
Therapy and Prevention
- antihistamine
Therapy and prevention
- washed rbcs
- IV fluid support (renal function)
- premedication
- RBC transfusion (anemia)
- for severe reactions: aminophylline, epinephrine,
- S&S of hemolysis and DIC should be monitored to
corticosteroids
reduce the risk of renal failure
- temporary cessation of blood component
- history of previous transfusion of pregnancy,
transfusion reaction
transplants and transfusion reaction

ANAPHYLACTIC & ANAPHYLACTOID REACTION

IMMEDIATE NON-HEMOLYTIC TRANSFUSION
REACTION → immediate hypersensitivity type of immune
response
→ occurs about 1% of transfusion reaction
→ anaphylactic can range from mild urticaria and
→ most commonly encountered
pruritus to severe shock and death
→ any 1°C or greater temp increase above the px → attributed to IgA deficiency (patients have
baseline during or w/in transfusion
developed anti-IgA from transfusion or pregnancy)
→ caused by leukocyte antibodies present in px
→ any organs can be involved: lungs, skin , blood
plasma vessels ,nerves, skin, gastrointestinal tract
→ Etiology: due to leukocyte antibodies
Distinguishing factors:
Signs & Symptoms - fevere is absent
• fever w/ or w/o chills - signs and symptoms occur after transfusion of just
• hypotension few milliliters of plasma or plasma containing blood
• cyanosis components
• tachycardia
• tachypnea Signs & Symptoms and Clinical Work-Up
• dyspnea • patient is deficient in IgA
• cough • sudden of onset with cough, dyspnea, nausea,
• limited fibrinolysis emesis, bronchospasm, flushing of the skin, chest
• transient leukemia pain, hypotension, abdominal cramps, diarrhea,
• history of transfusion reaction, transplantation, shock, loss of consciousness and death
pregnancy, drug therapy
ANAPHYLACTOID
Therapy and prevention → patients have normal levels of IgA but limited type
- leukocyte-reduced specific anti-IgA that reacts with light chain (kappa
or lambda) of the donor’s IgA
Malaika Ina M. Galos BSMLS-SP1 30
 → characterized by urticaria, periorbital swellling, → hypercalcemia leads to congestive heart failure and
dyspnea, perilaryngeal edema pulmonary edema
→ serum sample of patient can be
immunoelectrophoresed to determine IgA levels or Signs & Symptoms
immunodiffusion to identify subclass of antibody • dyspnea
• coughing
Therapy and prevention • orthopnea
- stop the transfusion and do not restart transfusion • chest discomfort
of blood component • headache
- IV lines should be given with NSS • restlessness
- give epinephrine immediately for severe reactions • tachycardia
- Corticosteroids or Aminophylline may be indicated • systolic hypertension
- vital signs must be stabilized • abnormal electrocardiogram results
- plasma should removed from the blood component
before transfusion Therapy and prevention
- transfuse blood compoment lacking IgA - transfusion should be stopped immediately
- slowest possible rate must be used
NON CARDIOGENIC PULMONARY REACTIONS - IV line should be maintained
(TRALI) - patient may be placed in a sitting position
→ Noncardiogenic Pulmonary Edema - ECG and chest x-ray to assess cardiac and
pulmonary status
→ Transfusion-Related Acute Lung Injury
- central venous pressure and vital signs should be
→ it is a pulmonary hypersensitivity reaction
monitored
→ Allergic pulmonary edema
- oxygen therapy and diuretics should be used
→ cause is not well understood
- therapeutic phlebotomy can be used
→ most consistent finding is Antileukocyte antibodies - cardiac arrythmia or decreased myocardial function
in donor or patient plasma (could initiate must be corrected
complement-mediated pulmonary endothelial injury) - transfusion rate should be 100 mL/hour or less
→ Antileukocyte antibodies could react with leukocytes - donor units should be split into aliquots to permit
to trigger the complement system to produce C3a transfusion to longer period
and C5a - washed red cells should be used to reduce plasma
→ this would cause tissue basophils and platelets to oncotic load
release histamine and serotonin, resulting in
leukocyte emboli aggregating in the lung capillary
bed BACTERIAL CONTAMINATION REACTION
→ low frequency,rapid onset and will lead to death
Signs & Symptoms and Clinical Work-Up → most are caused by Yersinia enterocolitica
• TRALI is characterzed by chills, cough, fever, → commonly caused by endotoxin produced by
cyanosis, hypotension, increasing respiratory bacterial grown in cold temperature
distress → Pseudomonas species, Escherichia, Yersinia
• mild resolving after a few days, severe rapidly enterocolitica
progressive pulmonary failure
• sera from donor and patient should be tested for Signs & Symptoms and Clinical Work-Up
antileukocyte antibodies • appear rapidly during transfusion or within about 30
mins. after transfusion
Therapy and prevention • reaction is termed as warm characterized by
- if signs and symptoms occur during transfusion, dryness and flushing of the skin
discontinue the transfusion • fever, hypotension, shaking , chills, muscle pain,
- Leukocyte-reduced blood component should be vomiting, abdominal cramps, bloody diarrhea,
used hemoglobinuria, shock, renal failure, DIC
- adequate respiratory and hemodynamic supportive • transfusion should be stopped immediately
treatment • IV lines must be kept open
• component unit, any associated fluid of the
TRANSFUSION ASSOCIATED CIRCULATORY transfusion equipment should be sent immediately
OVERLOAD to the blood bank for further inspection
• gram stain and culture
→ an iatrogenic (physician-caused) transfusion
• blood culture for the patient for detection of
→ few transfusion of a unit at too fast a rate
aerobic and anaerobic organisms
Malaika Ina M. Galos BSMLS-SP1 31
Therapy and prevention Signs & Symptoms and Clinical Work-Up
- broad spectrum antibiotics should be administered • maybe mild, slight fever and falling hemoglobin and
intravenously hematocrit levels
- therapy for shock, steroids (dopamine), fluid • severe, platelet refractoriness with bleeding
support, respiratory ventilation and maintenance of • if HLA antibodies are suspected, lymphocyte panels
renal function maybe indicated and lymphoctytotoxic antibody procedures are
- visual observation of RBC units for color change performed on patient’s serum
(brown/purple discoloration), visible clots or • thorough patient history of past transfusions,
hemolysis transplantations and pregnancies is important
- infusion of blood components within 4 hours
- adherence to policies and procedures regarding Therapy and prevention
blood component preparation, storage, handling - 3rd generation bedside leukocyte filters, delay if
and preparation not prevention of antileukocyte antibody production
- matching of donor and patient RBC phenotypes to
PHYSICAL & CHEMICALLY INDUCED avoid sensitization
TRANSFUSION REACTION
→ Heterogenous of transfusion reactions may include: POST-TRANSFUSION PURPURA
o physical RBC damage → Rare complication of blood transfusion, usually
o depletion and dilution of coagulation factors involving platelet concentrates
and platelets → Characterized by a rapid onset of thrombocytopenia
o hypothermia, citrate toxicity, hypokalemia or as a result of anamnestic production of platelet
hyperkalemia and air embolism alloantibodies
→ Can result in intravascular hemolysis → Usually occurs in multiparous females
→ 7-14 days lag time between transfusion and onset
Signs & Symptoms and Clinical Work-Up of thrombocytopeniia
• nonspecific, common are facial numbness, chills, → Platelet alloantibody attaches to the platelet
generalized numbness, muscle twitching, cardiac surface permitting extravascular destruction by the
arrythmias, nausea, vomiting, perioral tingling, RES in the liver and spleen
altered respirations and anxiety → Patient’s platelets are destroyed enhancing
• laboratory tests for electrolyte levels, calcium, blood thrombocytopenia
pH, glucose, U/A, Hb, Hct, PT and PTT
Signs & Symptoms and Clinical Work-Up
Therapy and prevention • Occur about 1-2 weeks after transfusion
- hypothermia, placing the patient in a warm blanket, • Thrombocytopenia can be severe <10,000/mm3
blood warmers should be used • Hematuria, melena, vaginal bleeding have been
- supportive care to any cardiac arrythmias or reported
electrolyte imbalance • Platelet transfusions for severe cases
- heparin might be indicated for DIC • Platelet count and coagulation support
- citrate toxity, administration of calcium-rich product
• Patient’s sera should be tested for platelet-specific
(milk, antacid with calcium gluconate)
antibodies, HLA antibodies and lyphocytotoxic
- monitoring patient’s mental status and vital signs
antibodies

DELAYED NONHEMOLYTIC TRANSFUSIO N Therapy and prevention

REACTION - Corticosteroids, exchange transfusions and
Alloimmunization plasmapheresis
→ may result from prior exposure to donor blood - Intravenous immunoglobulin therapy
components - Thorough patient history of prior transfusion and
→ adverse effect of blood transfusion any adverse reactions
→ lymphocyte memory is invoked, production of IgM
and IgG antibodies TRANSFUSION-ASSOCIATED GRAFT-VERSUS -
→ secondary exposure, production of large amounts of HOST DISEASE
IgG rising in the first 2 days after re-exposure to the → Complication of blood component therapy or bone
antigen marrow transplantation
→ antibody produced attaches to the antigenic surface → Death is usually caused by infection or hemorrhage
and may interact with the complement system or secondary to bone marrow aplasia
RES or MPS (monocyte phagocytic system)
Malaika Ina M. Galos BSMLS-SP1 32
 → Caused by proliferation of T-cell lymphocytes from
donor’s blood responding to major and minor Therapy and prevention
histocompatibility antigens in the patient - Removal of accumulated tissue iron stores
→ Patients with cell-mediated immunodeficiency are - Subcutaneous infusion of desferrioxamine (iron-
at risk of not being able to reject transfused chelating agent)
lymphocyte - Transfusion of few units of RBCs
→ Patient’s who have an HLA haploidentical type
donor (1st degree relatives) IMMUNOSUPRESSION
→ Generalized nonspecific effect that diminishes the
Risk groups:
activity of the recipient’s immune system after
- Patients experiencing lymphopenia or bone marrow
blood transfusion
suppression
→ Wide range of effects on transfusion recipient (for
- Fetus receiving exchange transfusion
further research)
- Individuals with congenital immunodeficiency
syndromes → No specific mechanisms have been definitely proved
- Patients with certain hematologic and oncologic → Several theories such as rapid uptake of blood
disorders component cellular matter into the RES or MPS
- Patients receiving blood components from blood
relatives TRANSFUSION REACTION INVESTIGATION
Necessary for:
Signs & Symptoms and Clinical Work-Up - Diagnosis
• Appear about 3-30 days post transfusion - Selection of appropriate therapy
• Pancytopenia, fever, elevated liver enzymes, - Transfusion management
copious, watery diarrhea, skin erythema - Prevention of future transfusion reactions
desquamation
• Tissue biopsies, lab tests for live function (liver, GTT, Should include:
skin, BM histologic changes) - Diagnosis
• Observation for infection or coagulation - Medical history of pregnancies, transplants and
abnormalities previous transfusions
• HLA typing to confirm the presence of donor’s cells - Current medications
in the patient’s circulation - Clinical signs and symptoms of the reaction

Therapy and prevention Immediate Laboratory Investigation

- Corticosteroids, cyclosporine, methotrexate, 1. Clerical checks (mislabeling and misidentification)
azathioprine and antithymocyte globulin - Patient & specimen identification
- Irradiated blood components - Blood unit inspection
- Tubing, filter and solution examination
IRON OVERLOAD - Label and record checks
2. Visual Inspection
→ Long term complication of RBC transfusion - Observe the serum of the patient pre and
→ Known as transfusion hemosiderosis post-transfusion reaction blood specimens
→ Each unit of RBC has about 225 mg iron - Color of pre and post-transfusion recipient
→ Patients chronically dependent on RBC transfusion serum/plasma for hemolysis
support 3. DAT-performed on post-transfusion specimen
→ Congenital hemolytic anemia 4. Additional testing
→ Aplastic anemia and chronic renal failure - ABO/Rh grouping
→ Accumulated iron affect the function of heart, liver - Compatibility test
and endocrine glands leading to hemosiderosis the - Ab & alloantibody identification
effect of the interference of mitochondrial function - Bilirubin
by excess iron accumulation - Hb & Hct
- Urine test
Signs & Symptoms and Clinical Work-Up
• Muscle weakness, fatigue, weight loss, mild MY PERSONAL NOTES FROM HENRY’S
jaundice, anemia, mild diabetes and cardiac FEBRILE NONHEMOLYTIC REACTIONS
arrythmias
• Assessment of storage iron level (ferritin) and other → Signs and symptoms: chills, rigors
iron studies should be performed → Defined as a rise in temperature of 1°C or greater
• Tissue stains for iron in tissue biopsies
Malaika Ina M. Galos BSMLS-SP1 33
 → Symptoms may be delayed for up to 1 hour after the → Subcutaneous epinephrine 0.3-0.5 mg (repeat that
procedure has been complete every 20 to 30 minutes for up to 3 doses)-
→ Self limited and usually resovles w/in 2-3 hours hypertension unresponsive to supportive measures
→ Premedications: Antipyretics (325-500mg), or significant bronchospasm
Dipenhydramine (25-50mg) → Intravenous epinephrine 0.5 mg (repeated every 5
→ The decision to restart the transfusion should be to 10 minutes)- refractory hypertension
driven by the 1clinical condition of the patient, 2the → Intravenous diphenhydramine 50-100 mg- urticaria
severity of the febrile reaction symptoms, 3the rarity → Aminophylline 6 mg- bronchospasm
of the blood product involved, and the 4results of → Patients with IgA deficiency who develop IgE anti-
transfusion reaction testing IgA can have anaphylactic reactions
→ Has been attributed to the accumulation of → Px who have significant allergic reactions should be
pyrogenic cytokines in units during storage evaluated for their quantitative IgA levels. Recent
(produced by leukocytes) transfusion may elevate serum IgA levels falsely
→ Prestorage leukocyte reduction- reduce the → Px with haptoglobin deficiency can have similar
generation of cytokines in platelets anaphylactic transfusion reactions due to IgG or IgE
→ Red blood cells may be more effective than PLR in antihaptoglobin
preventing febrile reactions → Haptoglobin deficiency is rare in North American
→ Febrile reactions to platelet transfusion may be populations but is more common than IgA deficiency
reduced by using platelet concentrates less than 4 among Japanese patients suffering anaphylactic
days old reactions

ALLERGIC REACTIONS ACUTE HEMOLYTIC REACTIONS

→ Signs and symptoms: pruritus, urticaria, erythema, → Signs and symptoms: fever(most sensitive initial sign
cutaneous flushing. Gastrointestinal involvement of AHTR) and chills, nausea, vomiting, pain,
may include nausea, vomiting, abdominal pain, dyspnea, tachycardia, hypotension, bleeding, and
diarrhea hemoglobinuria. During surgery, there is
→ May occur w/ any type of blood component, hypotension and excessive bleeding
including autologous RBCs → Lab findings: hemoglobinemia, hemoglobinuria,
→ Allergic reaction= transfusion should be elevated lactate dehydrogenase,
disconnected & IV access should be maintained hyperbilirubinemia, and low haptoglobin. Elevated
→ Upper airway involvment= prompt intubation BUN & Crea (if renal injury has occurred)
→ Dyspnea/ desaturation= oxygen administration → Present within 24 hours of transfusion
→ MILD allergy: dipenhydramine (50-100 mg) → Caused by transfusion of RBC units that are positive
→ SEVERE allergy: epinephrine for an antigen which the recipient has formed
→ REPEATED allergy: hydrocortisone (100 mg) antibodies. It can also be seen with transfusion of
→ Patients who have had repeated or significant antibodies (in donor plasma or intravenous
allergic reactions may benefit from PAS platelets or immunoglobulin [IVIg]) into recipients whose RBCs
from the concentration of cellular blood bear antigen recognized by those antibodies
components through the removal of most of the → Renal failure is a later complication
plasma or by the washing of red cells and platelets → Pain localizing to the flanks, back, abdomen, chest,
→ In mild cutaneous reactions, the transfusion may be head, infusion site
restarted after treatment without recurrence or → DAT (+) if the incompatible RBCs are present in the
worsening of the symptoms circulation
→ ABO incompatibility due to a clerical error is the
SEVERE ALLERGIC REACTIONS (ANAPHYLACTIC) most common cause of AHTRs
→ Signs and symptoms: cardiovascular instability → Transfusion should be discontinued and intravenous
including hypertension, tachycardia, loss of access should be maintained
consciousness, cardiac arrhythmia, shock, cardiac → identity of px and unit of RBCs should be
arrest. Respiratory involvement with dyspnea or reconfirmed
stridor → severity of AHTRs is related directly to the volume of
→ If anaphylactic reaction occurs the transfusion incompatible blood transfused
should be discontinued and IV access should be → Initial attention must be paid to cardiovascular
maintained support
→ Supportive care (intubation, oxygen, intravenous → Hypotension= fluid resuscitation & pressure
fluids, and placement of the patient in the support
Trendelenburg position), should be instituted
promptly
Malaika Ina M. Galos BSMLS-SP1 34
 → Maintenance of urine output with intravenous fluids → Antibiotic therapy should include broad- spectrum
and diuretics, mannitol or furosemide, early in the coverage, such as a β lactam and an aminoglycoside
course of the reaction has been used successfully → From the remaining blood component unit, a sample
→ An exchange transfusion with antigen-negative should be taken for Gram stain and culture. Blood
blood may be considered (with ABO incompatibility, cultures from the transfusion recipient should also
however, an exchange transfusion may greatly be collected at the time of reaction to compare with
reduce the chance of morbidity or death) the results of the unit culture
→ Septic transfusion reactions- frequent w/ platelet
DELAYED HEMOLYTIC REACTIONS transfusions
→ Methods (approved by the FDA):
→ Signs and symptoms: unexpected anemia, fever or
o culture, measurement of oxygen
chills, jaundice, pain, dyspnea, renal failure (rare),
consumption, and rapid qualitative
sickle crisis (SCD)
immunoassay
→ Lab findings: anemia, elevated lactate
o (Platelet PGD Test, Verax Biomedical
dehydrogenase, hyperbilirubinemia, low
Incorporated, Worcester, MA; BacTx Rapid
haptoglobin, leukocytosis, the presence of a new Assay System, Immunentics, Boston, MA) -
red cell antibody, DAT (+), elevated u. bilirubin closer to the time of issue and based on the
(active hemolysis)
immunological detection of
→ Occur at least 24 hours after transfusion lipopolysaccharide of gram (-) organisms
→ Most px present within the first 2 weeks after and teichoic acid of gram (+) organisms
receiving transfusion. Clinical DHTR may be → Apheresis platelets have a lower incidence of septic
recognized more than 6 weeks later reactions than pooled whole blood–derived
→ due to an anamnestic response to a red cell antigen platelets
to which the patient has previously made an → skin flora, if present, enter the collection at
antibody, the concentration of which was too low to venipuncture, diversion of the first 20 mL decreases
detect in pretransfusion testing. Rarely, a DHTR may the risk by 50%
be due to primary alloimmunization to a red cell
antigen
TRANSFUSION-RELATED ACUTE LUNG INJURY
→ Complications: renal failure or sickle crisis
→ Exchange transfusion= when there is a large burden → Symptoms: dyspnea, hypoxemia, tachycardia,
of Ag (+) red cells fever, hypotension, cyanosis. A chest x- ray usually
→ transfusion should be avoided until the causative shows pulmonary edema
antibody can be identified and antigen-negative units → Diff. diagnosis: circulatory overload, bacterial
obtained contamination, allergic reactions, acute respiratory
→ high-dose of IVIg = extravascular hemolysis distress syndrome (ARDS), pulmonary embolism,
→ single-dose of IVIg= prevent TR in alloimmunized px and pulmonary hemorrhage
whose compatible blood could not be obtained → Px with hematologic malignancy or cardiac disease -
high risk for TRALI
→ Resolves within 48-96 hours from outset
BACTERIAL CONTAMINATION OF BLOOD → A decrease in leukocyte or platelet count may be a
COMPONENTS useful clue in TRALI caused by transfusion of HLA
class I antibodies
→ Signs and symptoms: fever, chills, hypotension,
→ Mechanical ventilation= severely affected px
shock, nausea, vomiting, dyspnea, pain, diarrhea
→ TRALI has been attributed to the presence of
→ Complications result in shock, renal failure, DIC,
antibodies in the plasma of the transfused unit that
death
are directed against HLA or granulocyte antigens
→ Px factors: 1thrombocytopenia, 2pancytopenia present on recipient leukocytes. Some TRALI rxn,
→ Risk factors: 1contamination by gram(-) rods, may be due to the presence in the blood product of
2
greater px age, 3smaller volume of component lipid inflammatory mediators that activate already
transfused, 4younger age of stored platelet primed recipient neutrophils (from surgery, trauma,
concentrate or inflammation) to cause capillary injury and
→ RBCs contain: Acinetobacter, Escherichia, leakage
Staphylococcus, Yersinia, and Pseudomonas species → Plasma from multiparous female donors carry the
→ Platelet concentrates contain: greatest risk for HLA antibodies which can cause
o Gram (+) cocci: Staphylococcus, TRALI
Streptococcus
o Gram (-) rods: Acinetobacter, Klebsiella,
Salmonella, Escherichia, Serratia
o Gram (+) rods: Propionibacterium
Malaika Ina M. Galos BSMLS-SP1 35
TRANSFUSION-ASSOCIATED CIRCULATORY
OVERLOAD
→ Signs and symptoms: dyspnea, orthopnea, cyanosis,
tachycardia, elevated blood pressure, pulmonary
edema, jugular venous distention, pedal edema,
headache
→ Diff diagnosis: TRALI, allergic reactions, and causes
of congestive failure not related to transfusion, such
as valvular heart disease presents as congestive
heart failure during or shortly after transfusion
→ Px w/ pre-existing heart disease are at risk
→ Blood should be transfused slowly (transfusion
should be completed within 4 hours)
→ To avoid additional donor exposures, a unit can be
split using sterile technique and a portion retained in
the blood bank for later transfusion. Units can also
be concentrated by plasma removal HYPOTENSIVE REACTIONS
→ Absence of other transfusion reactions
GRAFT-VERSUS-HOST DISEASE → Occurs during transfusion
→ Signs and symptoms: rash, diarrhea, fever, liver → a drop of at least 10 mm Hg in systolic or diastolic
dysfunction, pancytopneia arterial blood pressure from the pretransfusion
→ Px w/ marked cellular immunodeficiencies- high baseline
risk → Hypotension begins during the transfusion and
o congenital cellular immunodeficiencies resolves quickly when the transfusion is
(DiGeorge syndrome, severe combined discontinued. If hypotension persists beyond 30
immunodeficiency syndrome) minutes after discontinuation of the transfusion,
o immaturity of the immune system another diagnosis should be strongly considered
(intrauterine transfusions, very- low- → Have been associated with red cell and platelet
birthweight infants) transfusions
o disease- associated immunodeficiencies → The patient should be positioned with head down
(Hodgkin lymphoma) and feet elevated (Trendelenburg position), and
o therapy- associated cellular isotonic fluids should be administered
immunodeficiencies (hematopoietic → Pressor support= if severe and refractory to IV fluids
progenitor cell transplantation, fludarabine → the condition is most likely due to the release of
treatment, antithymocyte globulin). bradykinin through activation of the contact
→ occur when viable donor T cells proliferate and are pathway of coagulation. Some reactions have been
not recognized by the recipient’s immune system as associated with angiotensin- converting enzyme
foreign but recognize and reject the host as foreign (ACE)–inhibitor drugs in the recipient and/or the use
→ has been reported with highly aggressive of leukocyte-reduction filters
chemotherapy for neuroblastoma, germ cell tumors o ACE is the major enzyme that breaks down
→ Patients with normal immunity may be at risk for bradykinin in the circulations
TA- GVHD if the donor is homozygous for an HLA
haplotype and the recipient is heterozygous but NONIMMUNE HEMOLYSIS
shares one haplotype (In this case, recipient
→ Signs and symptoms: hemoglobinemia,
lymphocytes are unable to recognize transfused
lymphocytes as foreign, but transfused cells see hemoglobinuria, Hyperkalemia (renal failure), Fever
recipient cells as foreign) → occur as a result of storage, handling, or transfusion
conditions
→ the bone marrow in TA- GVHD is of recipient type
and is attacked as well as the other organs → Hyperosmotic or hypo- osmotic fluids mixed with
red cells -> significant lysis
→ Exposure to ionizing radiation- cause chromosomal
damage & prevent replication of transfused → Transfusion of autologous blood from a patient
leukocytes (a midplane dose of 25 Gy, with a with sickle Hb -> hemolysis and death
minimum dose of 15 Gy is suggested) → All RBC units will show a slight degree in hemolysis
→ Licensed pathogen reduction system- alternative to with prolonged storage, but this will not result in
irradiation of platelets clinical signs after transfusion
→ Principal complications: renal failure & cardiac
arrhythmia (due to hyperkalemia)
Malaika Ina M. Galos BSMLS-SP1 36
 → Hemolytic transfusion reactions can rarely occur CYTOMEGALOVIRUS
when no red cell antibody is identifiable ▪ common in the general population
→ Serum K+ level should be checked ▪ In px with severe cellular deficiency, CMV causes
→ ECG- assess hyperkalemia pneumonitis, hepatitis, retinitis, multisystem organ
→ Urine output should be maintained with hydration failure
▪ 1
Seronegative & 2leukocyte reduced blood
TRANSFUSION-TRANSMITTED DISEASES components- minimize CMV transmission

PARVOVIRUS B-19
▪ Incidence of viremia among blood donors is variable,
with episodic peaks (1:5000)
▪ causes a mild self- limited febrile illness
▪ is trophic for erythrocyte precursors in the bone
marrow and can cause red cell aplasia,
HEPATITIS ▪ implicated in causing chronic anemia after bone
▪ HCV has accounted for the majority cases of marrow transplantation and in thalassemia
posttransfusion hepatitis
▪ PTH is asymptomatic in the majority of cases WEST NILE VIRUS
▪ HCV PTH eventually manifest clinical liver disease ▪ NYC- initial outbreak happpened
▪ PTH HBV is more severe ▪ Birds-reservoir; Mosquitoes-vector; Humans-
▪ If acquired before 5 years of age, acute HBV accidental host
infection is usually less severe, but the risk for ▪ acute WNV infections are asymptomatic (w/ febrile
chronic hepatitis is much greater illness & neuroinvasive disease in few infections)
▪ HDV requires coinfection for replication ▪ Nucleic Acid Testing- donor screening for WNV
▪ HBV & HDV- causes liver failure and hepatitis if
acquired simultaneously
▪ Transfusion-transmitted HDV is rare ZIKA VIRUS
▪ Screening tests for HBV is also effective for HDV ▪ Transmitted by Aedes spx. (sexual contact,
▪ HAV & HEV have relatively short periods of viremia transplacentally if pregnant)
and do not progress to a chronic carrier state ▪ Transfusion- transmitted Zika infection has been
▪ Hepatitis G virus (GBV- C) has a prevalence rate of documented in Brazil
about 1% in asymptomatic blood donors ▪ Licensed Nucleic Acid Testing- donor screening for
Zika Virus

HUMAN IMMUNODEFICIENCY VIRUS

▪ An acute viral syndrome occurs in about 60% of MALARIA
cases ▪ 1300 cases of malaria are reported annually in the
▪ The time to AIDS progression for transfusion US
recipients may be shorter than that for the ▪ TTD Malaria occurs rarely
implicated donor, but this may reflect the effects of ▪ donors are screened by a questionnaire detailing
age and other cofactors travel to endemic areas and symptomatology
▪ Group O HIV- not detected by EIA tests but endemic
in some areas like Western Africa [(resident= P. BABESIOSIS
deferral)- discontinued after the approval of ▪ Caused by Babesia microti (transmitted by ticks of
simultaneous combined assay] the genus Ixodes)- survives in refrigerated RBCs and
▪ combined immunoassay for simultaneous detection room temp platelet units
of HIV p24 antigen and HIV- 1 and HIV- 2 antibodies ▪ Many cases are asymptomatic and mild
that detects group O HIV was approved ▪ TTD Babesiosis is severe in asplenic or
immunosuppressed px
HUMAN T-CELL LYMPHOTROPIC ▪ Serologic and nucleic acid (required in high
▪ HTLV-I is associated with Adult T- cell prevalence areas) donor screening tests for B.
lymphoma/leukemia (ATL) and HTLV- associated microti
myelopathy (HAM)
▪ Most carriers are asymptomatic TRYPANOSOMA CRUZI
▪ HTLV-II is prevalent in IV drug users also associated ▪ Chagas disease
with HAM ▪ Risk is dependent on the demographics of donor
population
Malaika Ina M. Galos BSMLS-SP1 37
 ▪ Platelet transfusions causes majority of TTD T. Cruzi
infections
▪ T. cruzi is able to tolerate freeze-thaw cycles
▪ may be transmitted in plasma products as well as
cellular products

TRANSMISSABLE SPONGIFORM ENCEPHALITIS

▪ caused by a protein, called a prion (capable of
assuming an abnormal configuration resistant to
enzymatic degradation and serves as a template for
further abnormal prion deposition)
▪ vCJD- human TSE emerged from an epidemic of
bovine spongiform encephalitis (probably as a result
of entry of the pathogenic prion into the human food
supply from affected cattle)
▪ there is no available serologic test suitable for
testing donated units

Malaika Ina M. Galos BSMLS-SP1 38

(Lecturer: Sir Edgardo Monisit) 5. Equipment
6. Process management
7. Nonconformance management
QUALITY MANAGEMENT
8. Assessment
9. Continual improvement
Quality
- the degree to which the product service meets the
requirements Supplementary notes for blood donation
- provision of safe, satisfying donation experience for POST-DONATION CARE
donors
- accurately label & test blood components provided 1. No smoking for 2-3 hours after donation
to transfusion services 2. Drink more than the usual amount of fluids
- timely, accurate transfusion services provided to 3. Avoid strenuous activities for 24 hours especially
physicians and other health-care personnel lifting heavy objects
- safe & efficacious blood transfusion to patients 4. Do not drink alcohol within 24 hours
5. Leave the dressing or bandage for a minimum of 4
hours
BUILDING BLOCKS OF QUALITY 6. If you feel dizzy or lightheaded sit down with your
1. Quality Control head lowered between your knees and lie down
2. Quality Assurance with your feet elevated
3. Quality Management System (ex. AABB, Joint 7. Provide the telephone no. of the BSF (Blood Service
Commission International) Facility)
8. It is very important to thank the donor for his or her
blood donation
QUALITY CONTROL
▪ The daily testing for the reactivity of reagents,
POST-TEST NOTIFICATION
calibration of serologic centrifuge, monitoring of
1. Inform the donor that you would contact his or her
refrigerator, freezer and thawing devices
in case for any positive result
temperature
2. Send out letter (generic type)
▪ reveals a failure of an equipment system or a
process
▪ Activities include quality control of:
o collection instruments
o blood components
o reagents
o lab equipment

QUALITY ASSURANCE
▪ A set of planned actions that ensure that systems
and elements that influence the quality of a product
of service are working as expected, individually and
collectively
▪ tests how well the entire process is functioning
▪ Activities:
o measurement of the end product of
laboratory activity

QUALITY MANAGEMENT SYSTEM

▪ Provides framework for applying quality principles
and practices uniformly across all operations of a
department
▪ (a matter of what we do and what has to be done)

Essentials of QMS
1. Organization
2. Facilities and safety
3. Personnel
4. Purchasing and inventory
Malaika Ina M. Galos BSMLS-SP1 39
Malaika Ina M. Galos BSMLS-SP1 40
Malaika Ina M. Galos BSMLS| Adviser: Sir Edgardo Monisit

Immunohematology

ENZYMES THAT DESTROY AGS AND ABS FROM DIFFERENT BLOOD GROUP SYSTEMS
Le M&N S&s Fya and Fyb Jka and Jkb Anti- Lua Anti-Lub
Ficin  ✓ ✓ ✓  
Papain  ✓ ✓ ✓  
Bromelain ✓ ✓ ✓
Chymotrypsin  ✓ ✓ ✓
Trypsin ✓   ✓
Pronase ✓ ✓  ✓
DTT 
Glycine-EDTA 

BLOOD TYPE COMPATIBILITY

Blood Type Antigens Antibodies Can donate to Can receive from
A A B A,AB A,O
B B A B,AB B,O
AB AB X AB O,AB,A,B
O O AB O,A,B,AB O only

Malaika Ina M. Galos BSMLS- C3 41

BLOOD COMPONENTS & DERIVATIVES
Volume Shelf life
Whole blood 540 mL CPD – 21 days; CPDA1 – 35 days at 1-6 C
Red blood cells 260 mL (same as whole blood)
Red blood cells in additive solution 340 mL 42 days at 1-6 C
Washed RBCs 180 mL 24 hours at 1-6 C
Frozen Deglycerolized RBCs 250 mL 10 years at -65C to -200C & 24 hours at 1-6 C after washing
Leukocyte-reduced RBCs Similar to original unit Similar to original unit when closed system is used
2 units RBCs, Apheresis 320 mL 1 year at 1-6 C
Random Donor Platelet Concentrate 50-70 mL 5 days at 20-24 C with constant gentle agitation & for hours after pooling
Platelet, Pheresis 200-400 mL 5 days at 20-24 C with constant gentle agitation
Leukocyte reduced platelets Similar to original unit Similar to original unit when closed system is used
Granulocyte, pheresis 200-300 mL 20-24 C
Fresh Frozen Plasma 200-260 mL 1 year at -18 C or below
Cryopoor plasma 200 mL (same as FFP)
Cryoprecipitate 10-15 mL 1 year at -18 C or below & 6 hours after thawing
Factor VIII (Lyophilized) 25 mL 2 years at 2-8 C
Factor IX 25 mL 2 years at 2-8 C
Antithrombin III concentrate Per manufacturer’s directions 2 years at 2-8 C
S/D treated plasma 200 mL 1 year at -18 C
Donor-retested frozen plasma 200-260 mL 1 year at -18 C
Albumin 25-50 mL 5 years at 2-10 C
Plasma protein fraction 25-50 mL 3 years below 30 C
Immune globulin, Intravenous 10-2x0 mL 3 years at 2-8 C
Rh Immune globulin 1 mL 1 ½ years at 2-8 C

Malaika Ina M. Galos BSMLS-SP1 42

SIGNS AND SYMPTOMS (from Henry’s)
Febrile Allergic Severe Allergic Acute Hemolytic Delayed Bacterial Transfusion Transfusion Graft-Versus- Hypotensive Nonimmune
Nonhemolytic Transfusion Transfusion Transfusion Hemolytic Contamination Related Acute Associated Host-Disease Reactions Hemolysis
Transfusion Reaction Reaction Reaction Transfusion of Blood Lung Injury Circulatory
Reactions Reaction Components Overload
-chills -pruritus Cardiovascular -Fever!!! -unexpected -fever -fever -dyspnea -fever -Fever
-rigor -urticaria instability -chills anemia -chills -dyspnea -orthopnea -rash -hemoglobinemia
-erythema -hypertension -nausea -fever or chills -hypotension -hypoxemia -cyanosis -diarrhea -hemoglobinuria
-cutaneous -tachycardia -vomiting -jaundice -shock -tachycardia -tachycardia -liver -Hyperkalemia
flushing -loss of -pain -pain -nausea -hypotension -elevated blood dysfunction (renal failure)
consciousness -dyspnea -dyspnea -vomiting -cyanosis pressure -pancytopneia
Gastrointestinal -cardiac -tachycardia -renal failure -dyspnea -pulmonary
involvement arrhythmia -hypotension (rare) -pain A chest x- ray edema
-nausea -shock -bleeding -sickle crisis -diarrhea usually shows -jugular venous
-vomiting -cardiac arrest -hemoglobinuria (SCD) -pulmonary distention
-abdominal pain edema -pedal edema
-diarrhea Respiratory During surgery, -oliguria -headache
involvement -hypotension -DIC
-dyspnea or -excessive
stridor bleeding

SIGNS AND SYMPTOMS (from Henry’s)

IHTR DHTR INHTR ATR TRALI TACO Bacterial Physically & DNHTR PTP TA-GVHD Iron
Contamination Chemically Overload
Induced TR
-fever w/o chills -mild fever -fever w/ or -erythema -chills -dyspnea -fever -facial -fever -thrombocytopenia -pancytopenia - Muscle
-coagulopathy -moderate w/o chills -hives -cough -coughing -hypotension numbness -low hgb & hct -hematuria -fever weakness
-oliguria jaundice -hypotension -pruritus -fever -orthopnea -shaking -chills -platelet -melena -elevated liver -fatigue
-anuria -cyanosis -urticaria -cyanosis -chest discomfort -chills -generalized refractoriness -vaginal bleeding enzymes -weight
-death w/ Rare -tachycardia -hypotension -headache -muscle pain numbness -copious loss
sustained -oliguria -tachypnea -increasing -restlessness -vomiting -muscle -watery diarrhea -mild
-hypotension -DIC -dyspnea respiratory -tachycardia -abdominal cramps twitching -skin erythema jaundice
-cough distress -systolic -bloody diarrhea -cardiac desquamation -anemia
-limited hypertension -hemoglobinuria arrythmias -mild
fibrinolysis -abnormal -shock -nausea diabetes
-transient electrocardiogram -renal failure -vomiting -cardiac
leukemia results -DIC -perioral arrythmias
tingling
-altered
respirations
and anxiety

Malaika Ina M. Galos BSMLS-SP1 43

Malaika Ina M. Galos BSMLS| Adviser: Sir Edgardo Monisit

Immunohematology

Malaika Ina M. Galos BSMLS- C3 44

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