1

AUTOMATION IN
BLOOD BANKING

PRESENTER - Dr Shreya Prabhu

MODERATOR- Dr V Mahanthachar
WHY FULLY AUTOMATION IN 2

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

 3

 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
 4

 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.
 6

 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
 8

 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)
 9

 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)
 11

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.
 12

PRINCIPLE:
 Performed in a specially designed microtube.

 Based on controlled centrifugation of RBC through

a dextran-acrylamide gel that contains predisposed
reagents
13
 14

MICROTUBE
 15

GEL CARDS
16
17
 18

CENTRIFUGATION
19
 20
 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.
 21

 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 .
 22

 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.
 23

Photograph of agglutinated RBC’s trapped above the gel

matrix
25
 27

MIXED-FIELD REACTION
 28

 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
TESTS APPROVED BY FDA 29

 Gel technology is currently approved for

ABO forward and reverse grouping
Rh typing
DAT
Antibody screen
Antibody identification
Compatibility testing
 30

 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.
 32

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
 33

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
 34

SOLID-PHASE TECHNOLOGY
TESTS APPROVED BY FDA:

 Antibody screening

 Antibody identification

 Compatibility testing
 HISTORY 35

 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.
 SPRCA was developed by trade name Capture® 36
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
 37
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
 38
 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
 39

 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
 40

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
41
42
 43

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.
 44

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
 Solid phase technology for 45

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

EASE OF USE: 46

 Antigen are pre coated on test wells

 No pre-dilution of samples or reagents

 Same method and procedure for all assays

COST EFFICIENCY: 47

 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

 48
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 available Not available

ANTIBODY
SCREENING AND
CROSSMATCH
 49

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

 50

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 polystyrene
gels microplate wells

Testing detection AHG AHG Anti-IgG-coated red

cells
Washing required Yes No Yes

Centrifugation Yes Yes  Yes

 51
Reaction Quantitative: Quantitative: Semiquantitative
readings 1 to 4 1 to 4 g: strong
positive,
positive,
Negative, no MF

Stable reactions No yes (2–3 days) Yes (2 days)

Quality control Positive and Lot number of LISS color

negative control cards and diluent change, positive
on day of use and negative
control

Special No Yes Yes

equipment

Automation No Yes Yes

 52

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
 53

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
 54

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
 55

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
56
 57

 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
 59

GALILEO (IMMUCOR INC.)

60
 61

 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
 62

 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
 63
 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
 All reagents are bar-coded which helps in easy 64

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
Advantages 65

 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.
 Automation of Platelet Antibody screening and 66
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
 The system liquid used is 0.9% normal saline and 67
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
 68

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
 69

 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
 70

TECHNO (DIAMED)
71
 72

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
DISAVANTAGES 73

 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
 Instrument lacks the criteria of automated validation 74

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
 75

AUTOVUE
76
 77
 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
ADVANTAGES 78

 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

DISADVANTAGES 79

 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
 80
 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
AUTOMATED IDENTIFICATION 82

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.
 An eye readable number or description is included 83

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
 84

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
 85

 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
 86

 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
 87

MERITS DEMERITS
 Highly sensitive,  Expensive
specific  Maintenance
 Economical for large  Well trained man
blood banks power needed
 objective  Not ideal for small
blood banks
 89

THANK YOU 