Slide Presentation - Introduction To Immunohematology PDF

Introduction to Immunohematology
W. John Judd, FIBMS, MIBiol

Emeritus Professor
University of Michigan
Objectives
After reviewing this presentation you will be able to:
 Outline methods for detecting IgM and IgG antibodies

 Discuss the principle of gel technology
 Discuss the potential applications of blood group
genotyping
2 | Introduction to Immunohematology | W. John Judd, FIBMS, MIBiol | February 2012| Business Use Only
Immunohematology
 Pretransfusion testing
• ABO and Rh typing
• Antibody detection
• Crossmatching
 Investigation of immune hemolysis
 Perinatal testing
 Blood group phenotyping/genotyping
 Leukocyte and platelet serology
Antigens and Antibodies
BLOOD GROUP ANTIGENS:

Present (predominantly) on red blood cells
BLOOD GROUP ANTIBODIES

Present in plasma (or serum)
Antigen
An antigen is a substance (molecule) that, when

introduced into a human (or animal) who lacks
that substance, triggers the production of
antibody by the body’s immune system. The
antibody thus produced will react specifically with
the antigen in an observable way.
Blood Group Antigens
 324 blood group antigens recognized
 33 blood group systems
 40 unassigned antigens
 Molecular biology of assigned antigens is known
http5://www.isbtweb.org/fileadmin/user_upload/WP_on_Red_Cell_Immunogenetics
_and/Table_of_blood_group_antigens_within_systems_v2.0_110914.pdf
Antigen Function
ADHESION TRANSPORTERS
CARBOHYDRATES MOLECULES AND CHANNELS
ABO LW IN RH CO
H
P1PK XG SC RHAG GIL
I
LE FY MER2 JK XR
FORS
GLOB LU JMH DI
OK
COMPLEMENT STRUCTURAL OR
REGULATION ENZYMES UNKNOWN
CH/RG H KEL MNS

CROM I YT GE
KN FORS DO
System Notations
Common ISBT ISBT

Name Name Common Name Name
Rh RH P P1PK
Kell KEL Colton CO
Duffy FY Dombrock DO
Kidd JK Cartwright YT
Lewis LE MNS MNS
Diego DI Lutheran LU
Antigens, Genes and Phenotypes
System Antigens Genes Phenotypes
ABO A, B A, B, O A, B, O, AB
K-k+; K+k+; K+k- or

KEL K, k K, k
K-1,2; K1,2; K-1,2
Fy(a+b+); Fy(a+b-);
FY Fya, Fyb Fya, Fyb
Fy(a+b-)
P P1 P1, P2 P1, P2
Blood Group Antigens – Key Points
 Are present on RBCs as glycolipids, proteins or
glycoproteins
 Are inherited characteristics

 Have biological function
 Most are assigned to one of 31 blood group systems
Antibodies
An antibody, also known as an immunoglobulin, is

a protein produced by the immune system
following exposure to foreign antigen. Antibodies,
usually found in plasma, react with cells carrying
the foreign antigen in a very specific manner.
IgG Antibody
IgG vs. IgM Molecules
IgG vs. IgM Antibodies
Protein IgG IgM

Characteristic Immune Natural
Stimulus Protein Carbohydrate
Blood Group weight 150 kDa 900 kDa
Complement binding Rarely Yes
Antigen binding sites 2 10
Placental Transfer Yes No
Direct agglutinin Rarely Yes
Example Anti-Rh Anti-A, -B
Blood Group Antibodies
EXPECTED UNEXPECTED
 Natural anti-A  Alloimmune

 Natural anti-B  Autoimmune
 Passive
Blood Group Antibodies – Key Points
 Are stimulated by exposure to foreign antigens in the
environment, or by transfusion or pregnancy
 Are usually IgM and/or IgG immunoglobulins

 Anti-A and anti-B are expected antibodies (based on
RBC ABO type)
 All non-ABO antibodies are unexpected
Antigen-Antibody Reactions
Two Types of Tests
 Direct agglutination test for IgM antibodies

 Indirect antiglobulin test (IAT) for IgG
antibodies
Direct Tests – IgM Antibodies
ABO Typing
 Mix antibody and RBCs

 Incubate (optional)
 Centrifuge (1000 x g, 15 seconds)
 Examine
 Record results
IgM Agglutination
Agglutination
Indirect Antiglobulin Tests
For Detection of Immune (IgG) Antibodies

 Mix plasma (antibody) and RBCs
 Incubate at 37oC (10-60’)
 Centrifuge, examine, record (optional)
 Wash x 3 or 4 to remove unbound IgG
 Add antihuman globulin
 Centrifuge, examine, record
 Validate negative tests with IgG-coated RBCs
Detection of IgG Coating
Antihuman Globulin
Antigen-Antibody Reactions – Key Points
 Two types of tests are used to demonstrate blood
group antigen-antibody reactions
• IgM antibodies are used (or detected) by direct
agglutination tests
• IgG antibodies are used (or detected) by indirect
antiglobulin tests
 The indirect antiglobulin test (IAT) utilizes antihuman
globulin (AHG) reagent, otherwise known as Coombs
serum
Routine Serologic Tests
 ABO and Rh typing (Type)
 Detection of unexpected antibodies (Screen)
 Compatibility testing (Crossmatch)
 Phenotyping beyond A, B and D
 Direct antiglobulin tests (DAT)
Blood Bank Reagents – ABO
 Monoclonal anti-A and anti-B
 Anti-A colored blue, anti-B colored yellow to
confirm correct reagent has been added
 Pooled Rh-negative A1 and B RBCs

suspended in a preservative solution
ABO Typing
Traditional Tube Tests
RBCs Plasma
 2 tubes  2 tubes
• 1 drop anti-A  2-3 drops plasma to each
• 1 drop anti-B  1 drop 3-5% A1 RBCs
 1 drop 3-5% RBCs to  1 drop 3-5% B RBCs
each  Mix, centrifuge and
 Mix, centrifuge and examine
examine
ABO Typing
Expected Reactions
RBCs + Plasma +
Type Anti-A Anti-B A1 RBCs B RBCs
O 0 0 + +
A + 0 0 +
B 0 + + 0
AB + + 0 0
Anti-D for Rh Typing
TUBE TESTS:
Monoclonal IgM blended with either monoclonal
IgG or human (polyclonal) IgG in a low (6% wt/vol)
protein diluent
GEL MICROCOLUMNS:
IgM monoclonal anti-D
Rh Typing
DIRECT TEST TEST FOR WEAK D

 1 drop anti-D  Incubate negative direct tests
at 37oC
 1 drop 3-5% RBCs
 Mix, spin and read  Spin and read
 Wash x 4
 Add antihuman IgG
 Spin and read
 Confirm negative tests with
IgG-coated RBCs
ABO and Rh Typing – Gel
100 L plasma
10 L pRBCs +
50 L 0.8%
RBCs
RBC type A Plasma type

Anti-A Anti-B Anti-D Control A1 RBCs B RBCs
Distribution of ABO and Rh
O A B AB Rh+ Rh-
% US blacks 49 27 20 4 94 6
% US whites 45 40 11 4 83 17
% British 47 42 8 3 85 15
% Asians 33 30 27 10 99.5 0.5
ABO and Rh Typing – Key Points
 Can be done by tube, gel and solid-phase assays
 Two types of tests for RhD: a direct test, and an IAT to
detect weak expression of D
• Apparent D-negative donors (by direct tests) must be
tested for weak D
• Apparent D-negative patients need not be tested for
weak of D
 Different ethnic groups have disparate blood group
phenotype frequencies
Antibody Detection (Screening) –
AABB Requirements
 Clinically significant*
 37oC incubation ....... AHG
 No pooled RBCs
 Validate alternative methods
 IgG-coated RBCs
* Capable of causing a shortened RBC survival

Methods for Antibody Detection
Plasma:RBCs Time at 37°C AHG

SAL >2:1, 3-4% 30-60’ PS/IgG
ALB >2:1, 3-4% 15-30’ PS/IgG
LISS 2:2, 2% 10-15’ PS/IgG
GEL 1:2, 0.8% 15’ IgG
PEG 2:1, 3-4% 15-30’ IgG
LIP 2:1, 1% 1’ IgG
SPA 1:1, 0.4% 15’ IgG
Reagent RBCs – For Antibody Detection
 2-4 group O RBC samples that, between them, carry
C c D E e; K k; Fya Fyb; Jka Jkb; M N S s; Lea Leb and
P1 antigens
 DCe/DCe, DcE/DcE, and ce/ce
 Available commercially in preservative solution; new
shipment every 2-4 weeks
Two RBC Sample Set –
For Detecting Unexpected Antibodies
SCREEN D C c E e M N S s P1 Lea Leb K k Jka Jkb Fya Fyb

I R1R1 + + 0 0 + 0 + + + + + 0 0 + + 0 + + I
II R2R2 + 0 + + 0 + + 0 + + 0 + + + 0 + + + II
Factors Affecting the Sensitivity of Ag-Ab Reactions
 Plasma : RBC ratio  Antibody valency
 Time  Antibody concentration
 Temperature  Ionic strength
 pH  RBC surface charge
 Antigen-site density
Ionic Cloud
Steric hindrance of
antibody by ionic cloud
formed when RBCs are
suspended in saline
+-+-+-+-
LISS Tube Tests
2 drops serum Wash RBCs x 4

2 drops LISS
1 drop 3-5% RBCs
Add anti-IgG
10=15’ at 37oC Spin, read, grade
Confirm negative tests

Spin, read, grade with IgG-coated RBCs
Solid-Phase Assays
COAT WITH ANTIGEN ADD ANTIBODY
Y Y
+ AHG COATED RED BLOOD CELLS
Y Y
NEGATIVE REACTION POSITIVE REACTION

Gel Test
0.8% RBCs in LISS (50 L)
+ Sample (25 L)
Gel with Anti IgG
Incubated at 37oC for 15 min

Spin for 10 min
4+ 3+ 2+ 1+ 0
Anti-IgG
Gel Test Reaction
Antibody Detection – Key Points
 Popular methods for detecting unexpected antibodies
include LISS, gel and solid-phase adherence assays
 Automated platforms exist for both gel and solid-phase

methods
Antibody Identification
 Determining the blood group specificity of antibodies
causing a positive antibody screen
 Done by testing the plasma against a panel of reagent
RBCs of known phenotypes
Case Study
 63-year-old man
 Transfused 3 units of pRBCs after automobile
accident 3-years ago
 Scheduled for coronary bypass graft tomorrow –
2 units of pRBCs requested
Case Study – Initial Studies
RBCs + Plasma +
Anti-A Anti-B Anti-D Control A1 RBCs B RBCs
0 0 4+ 0 4+ 4+
SCREEN D C c E e M N S s P1 Lea Leb K k Jka Jkb Fya Fyb GEL

I R1R1 + + 0 0 + 0 + + + + + 0 0 + + 0 + + I 0
II R2R2 + 0 + + 0 + + 0 + + 0 + + + 0 + + + II 4+
Identification Panel
PANEL D C c E e M N S s P1 Lea Leb K k Jka Jkb Fya Fyb
1 r’r 0 + + 0 + + + + 0 0 0 + 0 + + + 0 + 1
2 R1R1 + + 0 0 + 0 + + + + + 0 0 + + + + + 2
3 R1R1 + + 0 0 + + 0 0 + + 0 + + + + + + 0 3
4 R2R2 + 0 + + 0 0 + 0 + + 0 + 0 + 0 + + + 4
5 r”r 0 0 + + + + 0 + 0 + 0 + 0 + + + 0 + 5
6 rrV 0 0 + 0 + + + 0 + + 0 0 0 + 0 + 0 + 6
7 rr 0 0 + 0 + 0 + 0 + + 0 + + 0 0 + + 0 7
8 rr 0 0 + 0 + + + + + + + 0 0 + + 0 + + 8
9 rr 0 0 + 0 + + 0 + 0 + 0 + + + + 0 0 + 9
10 rr 0 0 + 0 + + + + + 0 + 0 0 + + + + 0 10
11 Ror + 0 + 0 + + + 0 0 + 0 + 0 + + + 0 0 11
PATIENT AC
Findings
PANEL D C c E e M N S s P1 Lea Leb K k Jka Jkb Fya Fyb GEL
1 r’r 0 + + 0 + + + + 0 0 0 + 0 + + + 0 + 1 0
2 R1R1 + + 0 0 + 0 + + + + + 0 0 + + + + + 2 0
3 R1R1 + + 0 0 + + 0 0 + + 0 + + + + + + 0 3 0
4 R2R2 + 0 + + 0 0 + 0 + + 0 + 0 + 0 + + + 4 4+
5 r”r 0 0 + + + + 0 + 0 + 0 + 0 + + + 0 + 5 2+
6 rrV 0 0 + 0 + + + 0 + + 0 0 0 + 0 + 0 + 6 0
7 rr 0 0 + 0 + 0 + 0 + + 0 + + 0 0 + + 0 7 0
8 rr 0 0 + 0 + + + + + + + 0 0 + + 0 + + 8 0
9 rr 0 0 + 0 + + 0 + 0 + 0 + + + + 0 0 + 9 0
10 rr 0 0 + 0 + + + + + 0 + 0 0 + + + + 0 10 0
11 Ror + 0 + 0 + + + 0 0 + 0 + 0 + + + 0 0 11 0
PATIENT AC 0
Crossing Out – Cell #1
1 r’r 0 + + 0 + + + + 0 0 0 + 0 + + + 0 + 1 0
2 R1R1 + + 0 0 + 0 + + + + + 0 0 + + + + + 2 0
3 R1R1 + + 0 0 + + 0 0 + + 0 + + + + + + 0 3 0
4 R2R2 + 0 + + 0 0 + 0 + + 0 + 0 + 0 + + + 4 4+
5 r”r 0 0 + + + + 0 + 0 + 0 + 0 + + + 0 + 5 2+
6 rrV 0 0 + 0 + + + 0 + + 0 0 0 + 0 + 0 + 6 0
7 rr 0 0 + 0 + 0 + 0 + + 0 + + 0 0 + + 0 7 0
8 rr 0 0 + 0 + + + + + + + 0 0 + + 0 + + 8 0
9 rr 0 0 + 0 + + 0 + 0 + 0 + + + + 0 0 + 9 0
10 rr 0 0 + 0 + + + + + 0 + 0 0 + + + + 0 10 0
11 Ror + 0 + 0 + + + 0 0 + 0 + 0 + + + 0 0 11 0
PATIENT AC 0
1 r’r 0 + + 0 + + + + 0 0 0 + 0 + + + 0 + 1 0
2 R1R1 + + 0 0 + 0 + + + + + 0 0 + + + + + 2 0
3 R1R1 + + 0 0 + + 0 0 + + 0 + + + + + + 0 3 0
4 R2R2 + 0 + + 0 0 + 0 + + 0 + 0 + 0 + + + 4 4+
5 r”r 0 0 + + + + 0 + 0 + 0 + 0 + + + 0 + 5 2+
6 rrV 0 0 + 0 + + + 0 + + 0 0 0 + 0 + 0 + 6 0
7 rr 0 0 + 0 + 0 + 0 + + 0 + + 0 0 + + 0 7 0
8 rr 0 0 + 0 + + + + + + + 0 0 + + 0 + + 8 0
9 rr 0 0 + 0 + + 0 + 0 + 0 + + + + 0 0 + 9 0
10 rr 0 0 + 0 + + + + + 0 + 0 0 + + + + 0 10 0
11 Ror + 0 + 0 + + + 0 0 + 0 + 0 + + + 0 0 11 0
PATIENT AC 0
1 r’r 0 + + 0 + + + + 0 0 0 + 0 + + + 0 + 1 0
2 R1R1 + + 0 0 + 0 + + + + + 0 0 + + + + + 2 0
3 R1R1 + + 0 0 + + 0 0 + + 0 + + + + + + 0 3 0
4 R2R2 + 0 + + 0 0 + 0 + + 0 + 0 + 0 + + + 4 4+
5 r”r 0 0 + + + + 0 + 0 + 0 + 0 + + + 0 + 5 2+
6 rrV 0 0 + 0 + + + 0 + + 0 0 0 + 0 + 0 + 6 0
7 rr 0 0 + 0 + 0 + 0 + + 0 + + 0 0 + + 0 7 0
8 rr 0 0 + 0 + + + + + + + 0 0 + + 0 + + 8 0
9 rr 0 0 + 0 + + 0 + 0 + 0 + + + + 0 0 + 9 0
10 rr 0 0 + 0 + + + + + 0 + 0 0 + + + + 0 10 0
11 Ror + 0 + 0 + + + 0 0 + 0 + 0 + + + 0 0 11 0
PATIENT AC 0
Reaction Pattern
1 r’r 0 + + 0 + + + + 0 0 0 + 0 + + + 0 + 0 0
2 R1R1 + + 0 0 + 0 + + + + + 0 0 + + + + + + 0
3 R1R1 + + 0 0 + + 0 0 + + 0 + + + + + + 0 + 0
4 R2R2 + 0 + + 0 0 + 0 + + 0 + 0 + 0 + + + 4 4+
5 r”r 0 0 + + + + 0 + 0 + 0 + 0 + + + 0 + 5 2+
6 rrV 0 0 + 0 + + + 0 + + 0 0 0 + 0 + 0 + 6 0
7 rr 0 0 + 0 + 0 + 0 + + 0 + + 0 0 + + 0 7 0
8 rr 0 0 + 0 + + + + + + + 0 0 + + 0 + + 8 0
9 rr 0 0 + 0 + + 0 + 0 + 0 + + + + 0 0 + 9 0
10 rr 0 0 + 0 + + + + + 0 + 0 0 + + + + 0 10 0
11 Ror + 0 + 0 + + + 0 0 + 0 + 0 + + + 0 0 11 0
PATIENT 0 AC 0
Pretransfusion Testing – Goals
To provide blood and blood products for

transfusion in a timely, cost-efficient manner,
such that the transfused product provides
optimal clinical benefit to the recipient, but
does not cause adverse clinical effects or
transmit disease
Pretransfusion Testing
Donor/
Donor Patient Patient
History Requisition Selection
ABO/Rh Identification Crossmatch
Antibodies Sample Dispense

Disease ABO/Rh Bedside
ABO/Rh Antibodies
Records
Donor Testing – Blood Supplier
RBCs with anti-A and anti-B

ABO
Plasma with A1 and B RBCs
Direct tests with anti-D
Rh Confirm nonreactive units by a method that
detects weak expression of D
37°C incubation, IAT, may use pooled
Antibodies
RBCs
Infectious HBV, HCV, HIV, HTLV, WNV, Chagas,
Disease Syphilis, Bacteria
Transfusion Service Testing –
Donor Unit Confirmatory Tests
ABO Confirm all units (RBC type only)
Confirm Rh-negative units (direct

Rh
tests with anti-D)
Antibodies Not required
Infectious
Not required
Disease
Required Testing – Patient Samples
 RBCs with anti-A and anti-B

ABO
 Serum with A1 and B RBCs
 Direct tests with anti-D
Rh  Control system to recognize false-
positive tests
 37oC incubation
 Antiglobulin
Antibodies
 Clinically-significant
 No pooled RBCs
A Pint (Unit) of Blood
Blood Components – Derived from Whole Blood
 Packed Red Blood Cells (pRBCs)
 Plasma
• Fresh frozen – FFP
• Cryoprecipitate
 Platelets
 Granulocytes
The Crossmatch
 The final test that must be done before blood
is assigned to a patient
 Done to detect serological incompatibility
between donor RBCs and patient’s plasma
60
Selection of Components
Patient RBCs* Plasmaț

O O Any
A A, O A, AB
B B, O B, AB
AB Any AB
* Rh-negative RBCs for premenopausal females

ț Including Platelets and Cryoprecipitate
61
Crossmatch Methods
Unexpected Antibodies
Absent Immediate-Spin or Electronic
Present/History Indirect Antiglobulin Test
Other Important Tests
 Phenotyping, by direct agglutination or by the IAT for

antigens beyond A, B and D
 Direct antiglobulin test (DAT) on a patient’s RBCs (for

investigation of immune hemolysis)
Blood Group Genotyping –
Potential Research Applications
 Testing for rare donors
 Patients requiring chronic transfusions
 Patients with >1 antibody
 Patients with a positive DAT
 Recently transfused patients
 Antisera rare and unavailable
 Fetal RHD genotyping from maternal plasma
 Zygosity studies All Blood Group Genotyping tests commercially available in the U.S. and
Canada are for Research Use Only. Not for use in diagnostic procedures
Research Studies to Identify:
 Frequency of rare donors in geographic locations

 Frequency of multiple blood group systems in selected
donor populations
 Process improvement for advancing immunohematology

 Research to develop fundamental scientific knowledge
related to human disease conditions
All Blood Group Genotyping tests commercially available in the U.S. and
Patients Requiring Chronic Transfusions:
 Sickle-cell anemia patients

• African Americans
 Thalassemic patients
• Of Mediterranean origin, Arabs, Asians
 Match initial transfusions for Rh and K

 Complete match after first antibody made
 Patients with >1 Antibody:
• To predict additional antibodies that the patient may form
Patients with a positive DAT:
 Serologic phenotyping cannot be done readily by the

IAT
 Bound IgG can be removed by incubation with:

• Chloroquine diphosphate (time consuming; 80% effective)
• EDTA+glycine+HCl (denatures KEL antigens)
Recently Transfused Patients:
 Serologic phenotyping confounded by presence of

transfused RBCs
 Cell separation methods available

• High-speed microhematocrit centrifugation (tedious)
• Hypotonic saline (for sickle-cell disease patients)
Antisera Rare and Unavailable:
 Anti-k very rare

 Anti-Doa and anti-Dob are rare, impure, and require
use of enzyme-antiglobulin tests
 Predicting the extended phenotype of reagent

RBCs (DO, CO, YT, etc.)
Fetal RHD Genotyping from Maternal Plasma:
 Plasma from pregnant women contains soluble

fetal-DNA
 Done in Denmark and the UK to predict fetal Rh

type; RhIG withheld if Rh-negative
 Fetal DNA disappears soon after delivery

 SRY (male) and paternal VNTRs (female) used to
control for fetal DNA
Zygosity Studies:
 Paternal RHD zygosity determination

• RHD is flanked by two Rh boxes
• RHD is deleted in Rh-negative Caucasians, leading to
formation of a hybrid Rh box
 Zygosity determination of reagent RBC donors

• Dce/Dce vs Dce/ce
• Jka/Jka vs. Jka/Jk
Summary
 Elements of blood
 Antigens and antibodies
 Antigen-antibody interactions
 Pretransfusion testing
 Antibody identification
 Phenotyping
 Potential research applications of blood group genotyping

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Introduction to Immunohematology

W. John Judd, FIBMS, MIBiol

After reviewing this presentation you will be able to:

 Outline methods for detecting IgM and IgG antibodies

BLOOD GROUP ANTIGENS:

BLOOD GROUP ANTIBODIES

An antigen is a substance (molecule) that, when

CH/RG H KEL MNS

Common ISBT ISBT

System Antigens Genes Phenotypes

K-k+; K+k+; K+k- or

 Are inherited characteristics

An antibody, also known as an immunoglobulin, is

Protein IgG IgM

 Natural anti-A  Alloimmune

 Are usually IgM and/or IgG immunoglobulins

 All non-ABO antibodies are unexpected

 Direct agglutination test for IgM antibodies

 Mix antibody and RBCs

For Detection of Immune (IgG) Antibodies

 Pooled Rh-negative A1 and B RBCs

Type Anti-A Anti-B A1 RBCs B RBCs

DIRECT TEST TEST FOR WEAK D

RBC type A Plasma type

% Asians 33 30 27 10 99.5 0.5

* Capable of causing a shortened RBC survival

Plasma:RBCs Time at 37°C AHG

SCREEN D C c E e M N S s P1 Lea Leb K k Jka Jkb Fya Fyb

2 drops serum Wash RBCs x 4

10=15’ at 37oC Spin, read, grade

Confirm negative tests

COAT WITH ANTIGEN ADD ANTIBODY

+ AHG COATED RED BLOOD CELLS

NEGATIVE REACTION POSITIVE REACTION

Gel with Anti IgG

Incubated at 37oC for 15 min

Gel Test Reaction

 Automated platforms exist for both gel and solid-phase

Anti-A Anti-B Anti-D Control A1 RBCs B RBCs

SCREEN D C c E e M N S s P1 Lea Leb K k Jka Jkb Fya Fyb GEL

To provide blood and blood products for

ABO/Rh Identification Crossmatch

Antibodies Sample Dispense

RBCs with anti-A and anti-B

ABO Confirm all units (RBC type only)

Confirm Rh-negative units (direct

Antibodies Not required

 RBCs with anti-A and anti-B

Patient RBCs* Plasmaț

* Rh-negative RBCs for premenopausal females

Absent Immediate-Spin or Electronic

Present/History Indirect Antiglobulin Test

 Phenotyping, by direct agglutination or by the IAT for

 Direct antiglobulin test (DAT) on a patient’s RBCs (for

 Frequency of rare donors in geographic locations

 Process improvement for advancing immunohematology

Patients Requiring Chronic Transfusions:

 Sickle-cell anemia patients

 Match initial transfusions for Rh and K

 Serologic phenotyping cannot be done readily by the

 Bound IgG can be removed by incubation with:

 Serologic phenotyping confounded by presence of

 Cell separation methods available