Welcome Back to Immunohematology
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Transcript Welcome Back to Immunohematology
Identifying Antibodies
The Antibody Panel
CLS 422
Clinical Immunohematology I
Objectives
Discuss clinical situations when it is appropriate to
perform antibody identification.
Define a panel of cells.
Explain how the following factors aid in the
interpretation of antibody panels:
a.
Cross-out technique
b.
Variation in strengths of reaction
c.
Phases of reaction
d.
Autocontrol
e.
Red blood cell antigen typing
Objectives
List testing that can be performed to confirm
the identification of antibodies.
Identify the antibodies present, when given
panel results.
When is an antibody
identification panel
performed?
When the antibody screen is
positive.
When to perform test
The panel red blood cells (RBCs) are tested
against the patient’s serum or plasma in order
to identify the unexpected antibody or
antibodies present.
Identification
Antibody screen – positive
Run antibody panel to identify antibody (-ies).
If original panel does not provide a clear-cut
ID, test additional RBCs.
Selected cells
Alternate methods
Confirmation
Rule of 3 and 3
Antigen type patient’s RBCs.
Landsteiner’s Law!!!
The Panel
Series of 8 to 20 Group O RBCs
Various distribution of the most common RBC
antigens
Suspended in a preservative to protect antigen
integrity for 2 -4 weeks
Packaged with a lot-specific antigram
Antigram
Panel Antigram
Donor
Cell
number
D
RZR1
1
+
+
0
+
R1w
R1
2
+
+
0
R2R2
3
+
0
r’r
4
0
r’’r
5
rrK
Kp
Kp
Js
Js
Fy
Fy
Jk
Jk
Le
Le
a
b
a
b
a
b
a
b
a
b
+
0
+
0
+
0
0
0
+
0
+
+
+
0
+
0
+
0
+
+
+
0
0
0
+
0
+
0
+
+
+
0
+
+
0
0
+
0
+
0
+
0
+
0
+
+
0
0
+
0
+
0
+
+
0
+
0
+
0
+
+
0
+
0
+
0
0
+
0
+
0
0
+
0
+
0
+
+
0
+
0
+
0
0
+
0
+
+
9
0
0
+
0
+
0
+
+
0
+
R2r
10
+
0
+
+
+
0
+
0
0
R1R1
11
+
+
0
0
+
0
0
+
0
c
E
e
C
w
K
+
0
0
0
+
+
+
+
0
+
+
0
0
0
+
6
0
0
rrFya
7
0
Ror
8
rr
Patient
Cells
C
k
P
Lu
Lu
Xg
a
b
a
+
0
+
+
0
+
0
+
+
+
0
+
0
+
0
+
+
+
+
0
+
+
0
+
+
0
+
0
+
+
+
+
0
+
0
+
0
+
+
0
0
+
0
+
+
+
0
+
+
0
0
+
+
0
+
0
0
0
+
+
+
0
0
+
+
+
0
+
0
+
+
+
+
+
0
0
+
+
+
+
+
+
+
+
+
0
+
+
+
0
0
+
+
0
+
0
+
+
M
N
S
s
0
0
+
0
+
+
0
+
+
0
+
+
+
+
0
+
+
0
0
+
+
+
0
0
+
0
+
+
0
0
0
+
0
+
+
+
+
0
+
+
+
0
+
+
1
Auto Control
Patient’s serum/plasma tested against a
suspension of patient’s RBCs
Optional
Evaluate results in conjunction with patient history
Autoantibody
Newly forming alloantibody
If patient has a positive DAT, auto control will be
positive
Test Method
Usually the same as was used for the antibody
screen
Must include incubation at 37oC
Must include an AHG phase with reagent
containing anti-IgG
Interpreting Panel
Results
Cell
D
C
c
E
e
Cw
K
k
Kpa
Kpb
J
sa
J
sb
F
ya
F
yb
J
ka
J
kb
L
e
P
a
L
eb
M
N
S
s
L
ua
Lub
X
ga
A
H
G
1
CC
1
+
+
0
0
+
0
+
+
0
+
0
+
+
+
+
0
0
+
+
+
+
0
+
0
+
+
3+
2
+
+
0
0
+
+
0
+
0
+
0
+
0
+
0
+
0
+
+
+
+
0
+
0
+
+
0
2+
3
+
0
+
+
0
0
0
+
0
+
0
+
+
0
+
+
0
+
+
+
0
0
+
0
+
+
0
2+
4
+
0
+
0
+
0
0
+
0
+
+
0
0
0
+
0
0
+
+
0
+
0
0
0
+
+
0
2+
5
0
+
+
0
+
0
0
+
0
+
0
+
0
+
+
+
0
+
+
+
+
0
+
0
+
+
0
2+
6
0
0
+
+
+
0
0
+
0
+
0
+
+
+
+
+
+
0
+
0
+
0
+
0
+
+
0
2+
7
0
0
+
0
+
0
+
+
0
+
0
+
0
+
+
0
0
+
+
0
+
0
+
0
+
0
3+
8
0
0
+
0
+
0
0
+
0
+
0
+
+
0
+
+
+
0
0
+
0
0
+
+
+
0
0
2+
9
0
0
+
0
+
0
0
+
0
+
0
+
0
+
0
+
0
+
+
+
0
+
0
0
+
+
0
2+
10
+
+
+
0
+
0
+
+
0
+
0
+
+
+
0
+
0
0
+
+
0
+
+
0
+
+
3+
11
+
w +
+
w
0
0
+
0
+
0
+
0
0
+
0
0
+
+
0
+
+
0
0
+
+
0
2+
0
2+
Auto
Exclusion
Begin with the RBCs that failed to react
The antibody in the serum is not directed
against the antigens on these RBCs, so we can
eliminated these antibody specificities
Look at alleles to avoid problems with dosage!
Exclusion should be done using RBCs having
homozygous antigen expression.
Exceptions are low prevalence antigens
Exclusion
Cell
D
C
c
E
e
Cw
K
k
Kpa
Kpb
J
sa
J
sb
F
ya
F
yb
J
ka
J
kb
L
e
P
a
L
eb
M
N
S
s
L
ua
Lub
X
ga
A
H
G
1
CC
1
+
+
0
0
+
0
+
+
0
+
0
+
+
+
+
0
0
+
+
+
+
0
+
0
+
+
3+
2
+
+
0
0
+
+
0
+
0
+
0
+
0
+
0
+
0
+
+
+
+
0
+
0
+
+
0
2+
3
+
0
+
+
0
0
0
+
0
+
0
+
+
0
+
+
0
+
+
+
0
0
+
0
+
+
0
2+
4
+
0
+
0
+
0
0
+
0
+
+
0
0
0
+
0
0
+
+
0
+
0
0
0
+
+
0
2+
5
0
+
+
0
+
0
0
+
0
+
0
+
0
+
+
+
0
+
+
+
+
0
+
0
+
+
0
2+
6
0
0
+
+
+
0
0
+
0
+
0
+
+
+
+
+
+
0
+
0
+
0
+
0
+
+
0
2+
7
0
0
+
0
+
0
+
+
0
+
0
+
0
+
+
0
0
+
+
0
+
0
+
0
+
0
3+
8
0
0
+
0
+
0
0
+
0
+
0
+
+
0
+
+
+
0
0
+
0
0
+
+
+
0
0
2+
9
0
0
+
0
+
0
0
+
0
+
0
+
0
+
0
+
0
+
+
+
0
+
0
0
+
+
0
2+
10
+
+
+
0
+
0
+
+
0
+
0
+
+
+
0
+
0
0
+
+
0
+
+
0
+
+
3+
11
+
w +
+
w
0
0
+
0
+
0
+
0
0
+
0
0
+
+
0
+
+
0
0
+
+
0
2+
0
2+
Auto
Inclusion
Of the antibody specificities that have not been
excluded, match the pattern of positive and
negative reactions with the pattern of antigen
positive and antigen negative cells.
There must be an explanation for each positive
reaction seen.
Inclusion
Cell
D
C
c
E
e
Cw
K
k
Kpa
Kpb
J
sa
J
sb
F
ya
F
yb
J
ka
J
kb
L
e
P
a
L
eb
M
N
S
s
L
ua
Lub
X
ga
A
H
G
1
CC
1
+
+
0
0
+
0
+
+
0
+
0
+
+
+
+
0
0
+
+
+
+
0
+
0
+
+
3+
2
+
+
0
0
+
+
0
+
0
+
0
+
0
+
0
+
0
+
+
+
+
0
+
0
+
+
0
2+
3
+
0
+
+
0
0
0
+
0
+
0
+
+
0
+
+
0
+
+
+
0
0
+
0
+
+
0
2+
4
+
0
+
0
+
0
0
+
0
+
+
0
0
0
+
0
0
+
+
0
+
0
0
0
+
+
0
2+
5
0
+
+
0
+
0
0
+
0
+
0
+
0
+
+
+
0
+
+
+
+
0
+
0
+
+
0
2+
6
0
0
+
+
+
0
0
+
0
+
0
+
+
+
+
+
+
0
+
0
+
0
+
0
+
+
0
2+
7
0
0
+
0
+
0
+
+
0
+
0
+
0
+
+
0
0
+
+
0
+
0
+
0
+
0
3+
8
0
0
+
0
+
0
0
+
0
+
0
+
+
0
+
+
+
0
0
+
0
0
+
+
+
0
0
2+
9
0
0
+
0
+
0
0
+
0
+
0
+
0
+
0
+
0
+
+
+
0
+
0
0
+
+
0
2+
10
+
+
+
0
+
0
+
+
0
+
0
+
+
+
0
+
0
0
+
+
0
+
+
0
+
+
3+
11
+
w +
+
w
0
0
+
0
+
0
+
0
0
+
0
0
+
+
0
+
+
0
0
+
+
0
2+
0
2+
Auto
Other Points to Consider
In what phase(s) of testing is the antibody reactive?
Is the strength of reaction the same for each cell that
reacts, or is there variation in strength?
Dosage, antigen variability, or multiple antibodies.
Is the antibody reacting only with “homozygous”
cells of a certain specificity?
May give clue as to antibody identity and clinical
significance.
Weak antibody showing dosage.
Did the autologous control react?
Autoantibody or newly forming alloantibody.
Probability
Rule of 3 and 3
For each antibody specificity, are there 3
antigen positive cells that reacted and 3
antigen negative cells that did not react?
May use screen cells in addition to panel cells to
fill this rule
Cells do not need to have homozygous antigen
expression to fill this rule
The Rule of 3 and 3
Cell
D
C
c
E
e
Cw
K
k
Kpa
Kpb
J
sa
J
sb
F
ya
F
yb
J
ka
J
kb
L
e
P
a
L
eb
M
N
S
s
L
ua
Lub
X
ga
A
H
G
1
CC
1
+
+
0
0
+
0
+
+
0
+
0
+
+
+
+
0
0
+
+
+
+
0
+
0
+
+
3+
2
+
+
0
0
+
+
0
+
0
+
0
+
0
+
0
+
0
+
+
+
+
0
+
0
+
+
0
2+
3
+
0
+
+
0
0
0
+
0
+
0
+
+
0
+
+
0
+
+
+
0
0
+
0
+
+
0
2+
4
+
0
+
0
+
0
0
+
0
+
+
0
0
0
+
0
0
+
+
0
+
0
0
0
+
+
0
2+
5
0
+
+
0
+
0
0
+
0
+
0
+
0
+
+
+
0
+
+
+
+
0
+
0
+
+
0
2+
6
0
0
+
+
+
0
0
+
0
+
0
+
+
+
+
+
+
0
+
0
+
0
+
0
+
+
0
2+
7
0
0
+
0
+
0
+
+
0
+
0
+
0
+
+
0
0
+
+
0
+
0
+
0
+
0
3+
8
0
0
+
0
+
0
0
+
0
+
0
+
+
0
+
+
+
0
0
+
0
0
+
+
+
0
0
2+
9
0
0
+
0
+
0
0
+
0
+
0
+
0
+
0
+
0
+
+
+
0
+
0
0
+
+
0
2+
10
+
+
+
0
+
0
+
+
0
+
0
+
+
+
0
+
0
0
+
+
0
+
+
0
+
+
3+
11
+
w +
+
w
0
0
+
0
+
0
+
0
0
+
0
0
+
+
0
+
+
0
0
+
+
0
2+
0
2+
Auto
Antigen Typing
Confirms the antibody identification
Test patient’s RBCs (unknown antigen) against
appropriate anti-sera (known antibody)
Results should be
LANDSTEINER’S LAW
negative
Run positive and negative controls for anti-sera
A positive DAT or recent transfusion may invalidate
the typing results
Selecting Controls for Antigen Typing
Cell
D
C
c
E
e
Cw
K
k
Kpa
Kpb
J
sa
J
sb
F
ya
F
yb
J
ka
J
kb
L
e
P
a
L
eb
M
N
S
s
L
ua
Lub
X
ga
A
H
G
1
CC
1
+
+
0
0
+
0
+
+
0
+
0
+
+
+
+
0
0
+
+
+
+
0
+
0
+
+
3+
2
+
+
0
0
+
+
0
+
0
+
0
+
0
+
0
+
0
+
+
+
+
0
+
0
+
+
0
2+
3
+
0
+
+
0
0
0
+
0
+
0
+
+
0
+
+
0
+
+
+
0
0
+
0
+
+
0
2+
4
+
0
+
0
+
0
0
+
0
+
+
0
0
0
+
0
0
+
+
0
+
0
0
0
+
+
0
2+
5
0
+
+
0
+
0
0
+
0
+
0
+
0
+
+
+
0
+
+
+
+
0
+
0
+
+
0
2+
6
0
0
+
+
+
0
0
+
0
+
0
+
+
+
+
+
+
0
+
0
+
0
+
0
+
+
0
2+
7
0
0
+
0
+
0
+
+
0
+
0
+
0
+
+
0
0
+
+
0
+
0
+
0
+
0
3+
8
0
0
+
0
+
0
0
+
0
+
0
+
+
0
+
+
+
0
0
+
0
0
+
+
+
0
0
2+
9
0
0
+
0
+
0
0
+
0
+
0
+
0
+
0
+
0
+
+
+
0
+
0
0
+
+
0
2+
10
+
+
+
0
+
0
+
+
0
+
0
+
+
+
0
+
0
0
+
+
0
+
+
0
+
+
3+
11
+
w +
+
w
0
0
+
0
+
0
+
0
0
+
0
0
+
+
0
+
+
0
0
+
+
0
2+
0
2+
Auto
Value of Patient History
The following additional information may
assist in determining the identity of the
antibody:
History of antibodies
Transfusion, transplant, pregnancy (how many and
how long ago)
Medications
Diagnosis
Ethnicity
Reporting
Panel results are reported as “anti-” and then
the specificity
Anti-K
If specificity cannot be determined at this
point, additional testing must be performed
The End