Automation in blood banking

AUTOMATION IN
BLOOD BANKING
PRESENTER - Dr Shreya Prabhu
MODERATOR- Dr V Mahanthachar
1

WHY FULLY AUTOMATION IN
BLOOD BANK ?
To achieve “safe & timely blood transfusion”
 Faster TAT =>Faster blood transfusion
Reduce labour intensive manual test procedure
Test results directly upload to LIS (reduce
transcription error)
Thus, better quality of blood transfusion
2

Personnel shortages, TAT requirements and greater
regulatory demands have provided incentive for blood
services to seek automation
Automated equipments provide the level of quality
assurance required by new regulatory standards
Barcode reduces identification errors by providing
accurate patient identification
3

Automated systems from different manufactures
use advanced technologies like CAT (Column
Agglutination Technology) and SPRCA (Solid
Phase Red Cell Adherence)
Popular names for CAT and SPRCA are Gel and
Solid Phase technology respectively
Within the last several years automated methods
for typing blood, screening blood antibodies have
been developed.
4

The automated methods have led to a
remarkable improvement in blood safety
 The application of automation is now growing
in hospitals and transfusion services, because
human errors can lead to life threatening
consequences for the patients
6

Two basic Automation methodologies in blood
bank
I)GEL TECHNOLOGY (column agglutination using
a gel matrix)
II)SOLID PHASE TECHNOLOGY (solid-phase red
cell adherence)
8

The equipments that have been fully automated
and currently available in market are
Galileo (Immucor, Inc)- uses SPRCA
Techno (Diamed)- uses CAT
Auto Vue (Ortho Clinic Diagnostics Ltd)- uses CAT (beads)
9

GEL TECHNOLOGY
Gel technology is currently approved for ABO forward
and reverse grouping, Rh typing, DAT, antibody screen,
antibody identification and compatibility testing.
HISTORY:
In 1985,the gel test was developed by Dr Yves
Lappierre of France
Media used- Gelatin, acrylamide gel, glass beads
Gel appeared to be ideal material for trapping
agglutinates
Compared with traditional tube technology, the gel
test provides a more stable endpoint.
11

PRINCIPLE:
Performed in a specially designed microtube.
Based on controlled centrifugation of RBC through
a dextran-acrylamide gel that contains predisposed
reagents
12

Measured volumes of serum or plasma or RBCs
are dispensed into the reaction chamber of the
microtube
 The card is incubated and then centrifuged.
 The reaction chamber is actually a miniature test
tube, providing an area for the sensitization of
RBCs during incubation.
The shape and length of the column provides a
large surface area for prolonged contact of the
RBCs with the gel particles during centrifugation.
20

The gel particles are beads of dextran-acrylamide
that make up 75% of the gel-liquid mixture that is
preloaded into each microtube.
The gel particles are porous, they serve as a
reaction medium, filter, sieving the RBC
agglutinates according to size during
centrifugation.
Large agglutinates are trapped at the top of the gel
and are not allowed to travel through the gel during
the centrifugation of the card .
21

Agglutinated RBCs remain fixed or suspended in
the gel, while unagglutinated RBCs travel
unimpeded through the length of the microtube,
forming a pellet at the bottom following
centrifugation.
The gel test reactions are stable, allowing
observation for up to 3 days.
22

23
Photograph of agglutinated RBC’s trapped above the gel matrix

Negative reactions may appear mixed field when
incompletely clotted serum samples are used in
the gel test
Fibrin strands may trap unagglutinated RBCs
Before interpreting mixed-field, clinical history
should be considered
Ex- recently transfused or bone marrow transplant
recipients are expected to have mixed populations of
RBCs and commonly produce this
28

TESTS APPROVED BY FDA
Gel technology is currently approved for
ABO forward and reverse grouping
Rh typing
DAT
Antibody screen
Antibody identification
Compatibility testing
29

The ABO blood grouping card contains
gels that include anti-A, anti-B, and anti-A,B for
forward grouping.
Microtubes with buffered gel are used for ABO
reverse grouping.
The Rh typing card uses microtubes filled with gel
containing anti-D.
 Microtubes filled with gel containing anti-IgG are
used for compatibility testing, antibody detection,
and identification.
30

ADVANTAGES
Standardization is one of the major advantages,
(there is no tube shaking to resuspend the RBC
button)
It includes simple standardized procedures, no
wash steps.
Decreased sample volume needed for testing.
Provides stable, well defined end points of the
agglutination reaction
More objective, consistent and reproducible
interpretation of the test results
32

DISADVANTAGES
Need to purchase special incubators and
centrifuges to accommodate the microtube cards
used for testing.
A specific pipette must be used to dispense 25 mL
of plasma or serum and 50 micro L of a 0.8 percent
suspension of RBCs into the reaction chambers of
the microtubes.
Ideally suited to individuals who have been cross
trained to work in the blood bank
33

SOLID-PHASE TECHNOLOGY
TESTS APPROVED BY FDA:
Antibody screening
Antibody identification
Compatibility testing
34

 In 1978 Rosenfield and coworkers were the first to apply
the principle of solid-phase immunoassay to RBC typing
and antibody screening tests.
SPRCA (solid phase red cell capture assay) was
developed commercially for the detection of RBC and
platelet related antibodies.
In these test systems,one of the test reactants (either
antigen or antibody) is bound to a solid support (usually a
microtiter well) before the test is started.
The ability of plastics, such as polystyrene to absorb
proteins from solutions and bind them irreversibly made
SPRCA possible.
35
HISTORY

SPRCA was developed by trade name Capture®
by Immucor
In the Capture® immucor technology, tests were
adapted to microplate wells, either as full 96-well U-
bottomed plates or as 1x8 or 2x8 strips of U-
bottomed wells
Capture-R® for the detection of RBC antibodies and
Capture-P® for the detection of platelet antibodies.
To perform these tests, a laboratory centrifuge
capable of holding 96-well microplates is required,
along with a microplate incubator and an
illuminated reading surface or microplate reader
36

PRINCIPLE
Based on SPRCA
The tests uses chemically modified microplate test
wells in which intact reagent RBCs are bound to the
microwells before starting the test.
Patient serum or plasma and low ionic strength saline
are added to the RBC-coated microwells and
incubated at 370c to allow time possible for antibodies
to attach.
 After incubation, the wells are washed with pH-
buffered isotonic saline to remove unbound proteins
37

Wells are added with an indicator of anti-IgG–
coated RBCs, and the microwells are centrifuged.
Indicator cells are AHG coated RBCs
Centrifugation forces the indicator RBCs to
contact the immobilized sensitized reagent RBCs.
Positive test show adherence of indicator RBCs to
part or the entire well bottom, depending on the
strength of the reaction
38

In second generation tests, RBC membranes are
bound to the microplate test wells and air dried
Capture R® immucor is first generation solid
phase test, Capture R® Ready-Screen/Capture-
R® Ready-ID® are second generation tests
Solidscreen® is a solid phase test that detects
antibody using microplate wells that are coated
with polyspecific antihuman globulin
39

TEST REACTIONS
If antibody has attached to antigen , the indicator
cells will form a monolayer of RBCs
No antibody, nothing attached to antigen and
indicator cells will form a clearly delineated button
at the center of microplate well
Positive tests show adherence of indicator RBCs
to part or all of the well bottom, depending on the
strength of the reaction
40

ADVANTAGES
Standardization is the major advantage of solid-
phase technology.
Solid-phase technology provides stable, well-
defined endpoints of the reaction.
Ease of use
No predilution of reagents is required.
It is possible to test hemolyzed, lipemic, or icteric
samples.
The enhanced sensitivity makes the detection of
weak alloantibodies easier.
43

DISADVANTAGES
The major disadvantage is
the need for a centrifuge that can spin microplates,
a 37°C incubator for microplates
a light source for reading the final results.
Increased sensitivity may also be a disadvantage
in as much as solid phase may detect weak
autoantibodies that other systems miss
44

Solid phase technology for
immunohematology
 Capture has proved to be most dependable technology as it has
following features:
ACCURACY:
 IgG specific technology- detects clinically significant antibodies
 Sensitivity and specificity are higher
 Automated washing of hemolyzed, icteric, lipemic samples
assures validity
 Controls with each run assures validity
45

EASE OF USE:
Antigen are pre coated on test wells
No pre-dilution of samples or reagents
Same method and procedure for all assays
46

COST EFFICIENCY:
Higher lab productivity
Longer shelf life of coated antigens
Cost benefit of single antibody screen of patients
vs. number of crossmatches
Competitively priced
Easily automated or upgraded
47

PARAMETER ‘CAPTURE’ SPRCA OTHER TECHNOLOGIES
SENSITIVITY Highly for IgG antibodies Less
SPECIFICITY More False positivity is higher
SHELF LIFE 3-4 months 4-5 weeks
STANDARDIZATION Standardized Not
APPROVALS US FDA approved Non- US FDA approved
PLATELET
ANTIBODY
SCREENING AND
CROSSMATCH
available Not available
48

CURRENT TESTS AVAILABLE
Test Gel test Solid Phase
ABO- Forward Yes No
ABO- Reverse Yes No
Rh typing Yes No
Antibody screen Yes Yes
Crossmatch Yes Yes
Antibody identification Yes Yes
49

Traditional Gel Solid phase
Reaction chamber Tube Agglutination Microtube card Microplate wells
Reaction patterns Agglutination Agglutination Solid-phase
immune
adherence
Reaction matrix None dextran-acrylamide
gels
polystyrene
microplate wells
Testing detection AHG AHG Anti-IgG-coated red
cells
Washing required Yes No Yes
Centrifugation Yes Yes Yes
50

Reaction
readings
Quantitative:
1 to 4
Quantitative:
1 to 4
Semiquantitative
g: strong
positive,
positive,
Negative, no MF
Stable reactions No yes (2–3 days) Yes (2 days)
Quality control Positive and
negative control
Lot number of
cards and diluent
on day of use
LISS color
change, positive
and negative
control
Special
equipment
No Yes Yes
Automation No Yes Yes
51

SENSITIVITY
Technologies has been shown to detect at least as
many IgG Ab as conventional test tube methods
Sensitivity can be modified by pretreating the test
cells or monolayer with enzymes
Increased sensitivity is advantage when low titered
clinically significant antibody present
52

QUALITY CONTROL
GEL TEST
Uses special dispensers to prepare the RBC
suspensions and special pipettes to add measured
volume of plasma/serum
Each lot number of cards and diluent should be
tested on the day of use to confirm that the test
cards and diluted reagent RBC’s are reacting as
expected
53

SOLID-PHASE TECHNOLOGY
Recommends including a positive and a negative
control with each batch of tests
LISS formulated to detect the addition of plasma
by a color change from purple to blue when
plasma is added
54

AUTOMATION
Automated instruments use Gel and solid phase
technologies to increase the efficiency and
productivity of donor testing
They include ABS2000, ROSYS Plato(SPRCA),
Galileo,Diamed and ProVue by ortho clinical
diagnostics
These systems are capable of performing ABO/Rh
and antibody screens in the processing of donor
blood
55

The ABS2000 is credited as being the first fully
automated walk away system
It performs ABO/Rh using hemagglutination and
antibody screens/ crossmatches using solid phase
technology
The ABS2000 uses a bardode scanner to log
reagents and samples, transfer specimens by
automated pipetting, prepare RBC suspensions,
incubate, wash, centrifuge, read and interpret
results
Performs low to medium volume workloads
57

Galileo is a fully automated walk away system
which uses solid phase microplate technology to
perform all tests.
It uses SPARC technology for all coombs based
testing like
 AHG cross matching
antibody screening
antibody identification
DAT
For routine blood grouping testing, it uses
hemaglutination technology
61

Special and unique feature is availability and
automation of Platelet Antibody screening and
Platelet Cross matching
Manufactured by Immucor Inc, USA and used in
many countries
62

It has different components like
Centrifuge
Incubator
Washer
Camera
Sample and reagent loading bay
Microplate loading tower
which all are coordinated with each other to process
testing and no manual intervention is required
It has a sample capacity of 224 samples and
number of reagents can be loaded depending on
nature of tests performed
63

All reagents are bar-coded which helps in easy
identification of the same by instrument itself
There are various incubators of different
temperature available to carry different nature of
testing like blood grouping and antibody screening
simultaneously
Thus useful in institutions where different
parameters are performed simultaneously
Test results with images of microplates and all
other relevant details are continuously stored in
memory of instrument which can be reviewed as
and when required
64

Advantages
High loadging capacity- efficient to carry out heavy
workload
The capture technology used in Galileo is IgG
specific and hence provides high specificity
The capture technology used in Galileo is highly
sensitive
It has option to perform extended phenotyping for
all clinically significant antigens like Rh,Duffy,MNS
etc.
65

Automation of Platelet Antibody screening and
Platelet Cross matching makes it unique
The high processing speed of Galileo helps in high
through put of large batch testing within short time
The cell panels used for antibody screening and
identification in Galileo are in dried form coated on
microplate which increases the shelf life of these
panels for around 10-12 weeks
The washing step involved in assay procedure
enables the use of lipemic, icteric and hemolysed
samples
66

The system liquid used is 0.9% normal saline and
it can be replenished even during the test run
It has an option of online remote access through
internet for technical support and it can be used for
diagnostic purpose
Test results can be reviewed in four different
patterns by plate images, by grading of reactions,
by reaction strength only and by straight result as
positive and negative
The inbuilt quality control run as positive and
negative controls ensures validity of test results
67

Disadvantages
Closed system- no other reagents can be used other
than those by manufactures
If test run not managed properly, can lead to wastage
of consumables
Cost effectiveness must be considered as the
consumables are usually priced more
Internet connection is required for remote technical
support
68

The indicator cells used in coombs testing has
onboard shelf life of 24 hours
Archiving process must be done after regular
interval of 10-15 days to store all test results
69

ADVANTAGES
Has two working stations which function
independently and can be useful if one station is
not working
Gel technique is used is more sensitive
Interpretation of test results are done automatically
and image cards are available to review the results
The STAT position is available for urgent sample
processing
72

DISAVANTAGES
Not true continuous access and no sample can be
loaded during centrifugation of cards
Sample lodging capacity is of 36 samples
The blood grouping card used does not include
Anti-Ds from two different sources as
recommended by NACO, NBTC, BCSH and AABB
neither it has O cells in reverse grouping
The blood grouping card used also does not
include Anti-A,B which is essential for weak
subgroup of A group
73

Instrument lacks the criteria of automated validation
of test results as proper negative and positive
controls are not available during test run
Cards have to be manually placed- ? Fully automated
It is not possible to determine the presence of
clinically significant IgG antibodies when they are
present with cold antibodies as they mask the
presence of IgG reactivity in Gel Cards
The blood grouping profiles are fixed in gel format
and cannot be modified or changed according to
different needs of different blood banks
74

Uses Bead cards
ProVue is a walk-away instrument with capacity for
48 samples and 16 reagents
Safety features includes bar-code tracking system,
3 camera that record sample, reagent and card
identification
A camera in the instrument performs image
analysis and uses a mathematical algorithm to
interpret the results
77

ADVANTAGES
STAT Position is available for urgent testing of any
sample
Bead cards are sensitive
Partially continuous random access makes it better
in handling nature of testing simultaneously
Few reagents used
78

DISADVANTAGES
The Weak D testing not available
Loading capacity- 36
All samples scanned manually by hand barcode
scanner to feed samples in instrument
All samples centrifuged manually prior to loading
samples in instrument
79

The sample must be at least 1 ml for processing in
machine which is practically not possible in cross
matching for donor samples from tube segments
If any error occurs like waste container full, the
instrument stops working and waits for technician
to take required action
The machine produces many equivocal reactions
which must be edited manually by operator by
visual interpretation of cards seen on screen
80

AUTOMATED IDENTIFICATION
METHODS
BARCODES
A barcode comprises a series of vertical bars and
spaces arranged in various combinations to
represent different characters.
By combining the numbers, letters and other
characters, a series of barcodes can be built up to
represent donation numbers, blood groups and
various blood products
Barcodes have been very good for the blood bank
community. They offer high speed and accurate
data collection in a life-or-death application.
82

An eye readable number or description is included
with the machine- readable code.
Device which will interpret barcodes pass a beam
of light across the code making use of 2 levels of
optical reflectance viz. The black bars and white
spaces
83

ELISA
First test developed for the detection of
HIV antibodies in 1985 and currently widely
used test for serodiagnosis of HIV infection
Three types
Indirect ELISA
Competitive ELISA
Sandwich ELISA
Based on sandwich principle involving the use
of solid phase
84

Test sample is added to the microwell and
incubated
At the end of incubation period, the unbound
antibody/ antigen is removed by washing and
conjugate is added and incubated
Ag if present in the sample allows enzyme
conjugate to bind to the solid phase and build a
sandwich of Ab-Ag-Ab
Excess conjugate is washed away and chromogen
substrate is added for enzyme action to produce
color
85

The intensity of color is directly propotional to the
enzyme activity which in turn is propotional to the
Ag concentration in the sample or control
Result can be visualised or by ELISA reader
To determine reactive or non reactive important to
calculate the cut off value given by manufacturer
86

87
MERITS DEMERITS
Highly sensitive,
specific
Economical for large
blood banks
objective
Expensive
Maintenance
Well trained man
power needed
Not ideal for small
blood banks

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