Blood Banking
Blood Banking
Blood Banking
E. WEAK D (Du)
B. POSSIBLE GENE COMBINATIONS
1. Du is a weak form of D antigen rarely found among
Caucasians but common among in Blacks (22%).
Traditionally, Fisher-Race terminology is used to
2. Du red cells give weak or negative reactions with
describe Rh antigens whereas Weiner terminology is used
anti-D.
when describing Rh phenotype.
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e. Lewis antigens become weaker during b. Lewis antibodies are found in individuals who
pregnancy and females of Lewis-type Le (a-b+) have never been transfused or received other
during this period. antigenic stimuli.
c. Lewis antibodies are not known to cause
2. Lewis Pheontypes hemolytic disease of the newborn because of 2
a. There are 3 Lewis Phenotypes: reasons:
Le (a-b+) – present in secretors Most infants do not take up the soluble
Le (a+b-) – present in non-secretors Lewis antigen from the plasma to the red
Le (a-b-) – usually found in secretors cells by birth.
b. Rules regarding Le antigen expression Lewis antibodies are invariably IgM in form
A lele individual will not produce any and too large to cross the placental barrier.
antigen i.e. Le (a-b-).
A person who inherits at least one Le gene B. MNSs
at least one Se gene will be Leb positive i.e. 1. This was discovered in 1927 by Landsteiner and
Le (a-b+). Levine.
A person with at least one Le gene and 2. An individual is either MM, MN or NN.
sese genes will be Le (a+b-). 3. MNSs system demonstrated dosage effect i.e. MM
or NN reacts stronger than MN with anti-M and
Adult anti-N.
Levels 4. M and N are co-dominant alleles in paternity
Genes Reaction testing. The antigens are fully developed at birth.
Phenotype Secretors 5. The MNSs system differs from the ABO antigen in
inherited with
anti-Lea three ways:
- Leb a. Antigens found on red cells and some tissues
Le, H, Se Le (a-b+) 0 + A, B H Lea but not secretions.
70% and Leb b. Anti-M and anti-N are usually a result of a
substances transfusion or pregnancy.
Le, H, se Le (a+b-) + 0 Only Lea c. Anti-M and anti-N rarely occur naturally.
20% substances
Le, H, Se Le (a-b-) 0 0 10% of le 6. MNS antibodies
10% (a-b-) are a. Anti-M and anti-N are IgM and react best at 4C.
secretors b. The clinical importance of this system is in
Le, H, se Le (a-b-) 0 0 Transfusion reactions due to anti-S and
10% anti-s.
Medico legal cases (paternity)
c. Se controls ABH secretions, but has no control HDN (due to anti-S, anti-s and anti-U)
over Le secretion.
7. Problem (Paternity Testing)
3. Lewis antibodies Father: MM
a. At least four antibodies can be differentiated Mother: NN
Anti-Lea is most commonly found. It is Baby: MN
usally accompanied by anti-Leb especially Question: Is the alleged father excluded
in Blacks possessing Lewis type Le (a-b-).
Anti-Leb exists in two forms: anti-Lebh and Reactions:
anti-Lebl. The difference are as follows: Anti-M Anti-N
Cells Reactions Father MM 4+ Negative
O Le(b+) III Mother NN Negative 3+
A 2 Le(b+) III Baby MN 3+ 3+
Anti-Lebh
A1 Le(b+) o/w
H substance Neutralized Answer: The alleged father is not excluded.
Cells Reactions
Anti-Lebl O Le(b+) III
A2 Le(b+) III
A1 Le(b+) o/w
H substance Neutralized
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D. P SYSTEMS 2. Kx substance
1. The antigens formed are P1, Pk and P. a. Kx (precursor substance of Kell antigens) is
2. The P1 antigen is present on the red cells of 79% of present on the WBC and RBC of most
the population. Individuals who lack P1 are termed individuals.
P2. b. Kx is lacking from red cells, the cells have an
3. Individuals who lack P1P1 or Pk antigens are termed abnormal shape (acanthocytes) and a reduced
p. in vivo survival. Such individuals are said to
have McLeod phenotype.
4. Antibodies c. The absence of Kx from leukocyte has been
a. Anti-P1 is an IgM and has never caused HDN described by individuals with chronic
but has occasionally caused a transfusion granulomatous disease. The leukocytes are
reaction. In transfusion, recipients with anti-P1 able to phagocytose but not kill bacteria.
must be given P1 negative units. Patients with CGD have recurrent bacterial
b. Anti-P has the specificity of the biphasic infection.
Donath-Landsteiner antibody present in
Paroxysmal Cold Hemoglobinuria. 3. Kell antibodies
a. Kell antibodies are IgG and are stimulated by
E. LUTHERAN BLOOD GROUP SYSTEM transfusion or pregnancy. They can cause HDN
1. The antibodies usually associated with this system and HTR.
are:
a. The anti-Lua is a rare IgM antibody which does H. THE DUFFY (Fy) BLOOD GROUP SYSTEM
not cause HDN and HTR. 1. Duffy Antigens
b. Anti-Lub is an IgA which may cause HDN and a. The most important antigens are Fya and Fyb.
HTR. Fya is more immunogenic than Fyb.
c. Anti-LuaLub is IgM or IgA, and is active at 37C in b. The four phenotypes are:
AHG or enzyme. Fy (a+b+)
Fy (a+b-)
Fy (a-b+)
Fy (a-b-)
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Red cells that are Fy (a-b-) are resistant to 4. Cartwright Yta, Ytb Yta is poorly
penetration from Plasmodium vivax and developed at birth
Plasmodium knowlesi but Fy (a+) or Fy (b+)
cells are susceptible to this parasite since Yta is more
they invade the red cells at the Duffy common in
antigens. Chinese, Japanese
and North
2. Duffy Antibodies American
a. Most antibodies are IgG and can cause HTR 5. Diego Dia, Dib
and HDN. 6. Scianna Sm (Sc: 1)
Bua (Sc: 2)
I. KIDD BLOOD GROUP SYSTEM 7. Dombrock Doa, Dob
1. Kidd Antigens 8. Colton Coa, Cob
a. The antigen Jka and Jkb are weakly 9. Others
immunogenic.
b. The phenotypes are: a. Chido Cha Found on the C4d
Jk (a+b-) fragment of
Jk (a+b+) complement
Jk (a-b+)
Jk (a-b-) This is more commonly found in b. Rodgers Rga Adsorbed onto the
Orientals particularly Filipinos. RBC membrane
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Chapter 9: Antibody Detection & Identification 4. Requirements for Antibody Detection and
Identification
A. UNEXPECTED ANTIBODIES a. Reagent Red Cells (Antibody Screening Cells)
1. Clinically unexpected antibodies occur in less than Reagent red cells are commercially
3% of the population. Multiple antibodies are more prepared Group O red cell suspensions
frequently seen in: obtained from individual donors that are
a. Women than men because of possible phenotyped for the most commonly
sensitization during pregnancy. encountered and clinically important RBC
b. Among patients older than 60 years old who antigens.
have undergone transfusion multiple times. Group O cells are used so that naturally
occurring expected anti-A or anti-B will not
B. ANTIBODY SCREEN interfere with detection of unexpected
1. Defintion antibodies.
Screening cells are detected so that the
Antibody screen involves the reaction between following antigens are present on at least
patient serum or plasma with 2 or 3 reagent one of the RBC samples.
phenotyped for multiple antigens. It is also called
antihuman globulin test. D, C, E, c, e, M, N, S, s, P1, Lea, Leb, K, k, Fya,
Fyb, Jka, Jkb
2. Purpose of Antibody Screen
The reagent red cells are provided in either
The purpose of antibody screen is to detect red two or three vials. Each vial contains cells
blood cell antibodies other than expected anti-A from a single, different donor. The vials are
and anti-B antibodies. These antibodies are called provided with a description of the antigen
“unexpected antibodies” since they are present content of each of the cell. This description
only in 0.3 to 2% of the general population. is provided in a chart known as an
antigram.
3. Importance of Antibody Screen
a. Selection of appropriate blood for transfusion b. Enhancement Reagents
b. Investigation of HDN, immune hemolytic Enhancement reagents are solutions added
anemia, and transfusion reactions. to serum and cell mixtures to promote
antigen-antibody binding or agglutination.
C. ANTIBODY IDENTIFICATION The enhancement reagents are:
1. If the antibody screen is positive, and antibodies o Low ionic strength saline (LISS)
are present in the serum, a more definitive test o Polyethylene glycol (PEG)
called antibody identification or panel is o Bovine albumin
performed.
2. Another test to detect and identify antibodies is c. Antihuman globulin reagents (AHG)
the direct antiglobulin test (DAT). AHG is used to promote agglutination of
DAT is a test to detect antibodies coating the RBC sensitized with immunoglobulin G
surface of red cells in vivo. An antibody (IgG) or complement molecules.
detection method or panel also identifies these
antibodies. d. Coombs Control Red Blood Cells
These antibodies may be removed from red Coombs Control cells are RBC coated with
cells by process known as elution and then human IgG. They are prepared by
identified by a panel. incubating D positive RBC with potent anti-
3. Flow Chart of Antibody Identification D.
They are used to ensure that AHG tests
with negative results are not false
negatives because of the inactivation of
the AHG reagent.
In a negative AHG test, there is a free AHG
reagent in the test tube. When Coombs
control cells are added, the free AHG in the
test should cause agglutination of the
sensitized RBC.
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Typing of Immunoglobulin
o The typing of immunoglobulin is
accompanied by the use of
dithiothreitol (DTT) and 2-
mercaptoethanol (2-ME).
o DTT and 2-ME are used to remove
or inactivate IgM, leaving the IgG
intact. They are sometimes useful
to remove or break up red cell
agglutination by strong IgM cold
autoantibodies.
Enzyme Testing
o Enzyme testing is done to facilitate
the agglutination of red cells in the
presence of certain antibodies.
o In the procedure, RBC are exposed
to proteolytic (or protein-digesting
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