Recent advances in blood banking include the introduction of gel technology for blood grouping and cross matching. This provides more sensitive, specific, and efficient testing compared to traditional methods. Automation has also increased at various levels of blood banking through use of systems like the automated immunohematology testing system ORTHO AUTOVUE INNOVA. Additionally, new technologies like apheresis allow for collection of individual blood components from a single donor rather than whole blood, reducing donor exposure and providing purer products.

1. RECENT ADVANCES IN
BLOOD BANKING
By: Dr Vinanti Golwilkar
Guide: Dr K.K.Kulkarni

2. • Introduction
• Historical aspects
• Indian Scenario
• Recent advances in transfusion medicine with respect to:
-Blood collection & storage
-Grouping & cross-matching
-TTD testing
-Blood component separation: Apheresis
• Blood substitutes
• New technologies in blood banking
• Automation in blood banking
• Hemopoietic stem cell transplant
• Cord blood banking
• References

3. Introduction
• Transfusion Medicine has emerged as a separate
branch with the rapid & numerous advances in the field
of blood banking
• Notable among recent advances is gel technology for
blood grouping & cross matching, the technique of
apheresis, automation at various levels, introduction of
peripheral blood stem cell & cord blood banking

4. Historic aspects
• Ancient Egyptians bathed in blood and aristocrats
consumed blood so as to cure disease.
• 1616:Concept of circulation of blood by Sir William
Harvey
• 1665: First canine transfusion: Royal Society of London
• 1667: Animal to human transfusion: J Denis
• 1818:Human to human transfusion: James Blundell
• 1869:Use of nontoxic anticoagulant- sodium phosphate:
Braxton Hicks
• 1901:Discovery of ABO blood groups and transfusion
reactions : Nobel prize awarded to Karl Landsteiner

5. • 1916:The first anticoagulant preservative (citrate-glucose)
was used to preserve rabbit blood for 2 weeks.
• Rous-Turner’s solution was used for storage of blood
during the First World War.
• 1936: First Hospital Blood Bank in the United States
established at the Cook County Hospital in Chicago
• 1939:Discovery of hemolytic disease of newborn: Levine
and Stetson.
• 1940: Discovery of Rh factor and anti Rh: Landsteiner and
Weiner
• 1940: Discovery of human antibody similar to antiRhesus :
Weiner and Peters
• 1940: Dr. Charles Drew researched & documented a
technique for long term preservation of plasma

6. •1940: Edwin Cohn developed cold ethanol fractionation, a
process of breaking down blood plasma into components &
products
World War II stimulated blood preservation research and
developed techniques in blood transfusion.
The pioneer work of Dr. Charles Drew during World War II led
to the establishment of a world wide system of blood banks
•1950: Freeze drying of plasma - New concept of component
therapy
•Introduction of the plastic bag for blood collection & storage
•1959: First attempt of Bone Marrow Transplant in France

7. •1965: Judith Graham Pool developed technique known
as cryoprecipitation for concentrating factor VIII from
blood plasma
•1972: The FDA Bureau of Biologics began regulating
blood resources
•1977: The formal FDA compliance program was
established
•1978: CPDA-1 was developed, ↑ shelf life of RBCs for 35
days
•1988: First umbilical cord blood transplant
•1999: NAT for HIV and HCV

8. Indian Scenario:
Advances over the last decade
• 1999: Unsafe blood & blood products accounted for >9%
of AIDS cases
• 2004-2005: Unsafe blood & blood products accounted
for 2.07% of AIDS cases
• Ensuring widespread availability of safe & quality blood
is a critical component of National AIDS Prevention &
Control Programme under National AIDS Control
Organisation (NACO)

9. Phase I (1992-1999):
• Infrastructure development
• The National & State Blood Transfusion Councils(SBTC)
were registered as societies in 1996
Phase II (1999-2006):
• Expanding & building upon blood safety activities carried
out in Phase I.
• Introduction of new programmes like Accreditation of
Blood Banks, External Quality Assessment Scheme
(EQAS) for HIV testing, organizing workshops for
clinicians
• April 2002: A National Blood Policy formulated by NACO
was adopted by Govt. of India.
• July 2003: A meticulous action plan on blood safety was
finalised & adopted by the Govt. of India.

10. • Proportion of voluntary blood donation increased from
20% at start of phase II to present figure of >50%.
Phase III (2007-2012):
Goals:
• Regular voluntary non-remunerated blood donors should
form the main source of blood through phased increase
in donor recruitment & retention
• Promote appropriate use of blood, blood components &
blood products
• Aim to reduce transfusion associated HIV transmission
to <0.5%
• Develop long-term policy for capacity building to achieve
efficient & self sufficient blood transfusion services

11. Strategies:
1. Establish proper institutional mechanism for planning &
implementation of blood safety:
• Blood storage centres – Regional blood transfusion
centres – State Blood Transfusion Council – National
Blood Transfusion council
• Blood Bank Cell should be established in the office of
DCG(I) & State Drug Control Authority
2. Encourage rational blood use for transfusion:
Component separation facility should be set up at all
tertiary care institutions & at least 50% of the collection
should be separated into components

12. 3. Steadily increase the proportion of voluntary blood
donation upto 80% of the blood units collected, 50% of
them being from regular voluntary blood donors
4. Enhance & institutionalise Quality Assurance in Blood
Banks & also increase the counselling to those
voluntary blood donors whose samples test reactive

13. Blood collection
• Blood donation:
Paid or professional donors – Family or replacement
donors – Voluntary non-remunerated donors
Professional blood donation is banned in our country w.e.f.
1st
January 1998.
• Autologous blood donation:
-Blood collected from a patient for re-transfusion at a later
time into the same individual is called autologous blood.
-Safest possible blood for the patient

14. • Anticoagulant commonly used:
CPDA-1: Storage of blood upto 35 days
• Additive systems: To extend the red cell storage to 42
days & to harvest maximum amount of plasma, additive
systems (eg: Adenine Saline) are now available in which
the storage environment of red cells is altered by adding
certain nutrients after removal of plasma
Blood Storage

15. Grouping & Crossmatching
• Traditional methods:- Slide or tile method
- Tube method
- Microplate method
• Newer methods:- I.D.Micro-typing system
(Gel Technology)
-Glass microbeads technology
-Erythrocytes Magnetized technology
(Automated or Semi-automated instrumentation)

16. Gel Technology
• Innovative approach to red cell serology.
• Invented by Dr.Yves LapierreDr.Yves Lapierre of France in 1988.
• Developed to minimize problems associated with
conventional techniques of blood grouping.
• Addresses the issues of standardization and
documentation with unmatched sensitivity, specificity
and efficiency.
• DiaMed-ID Microtyping system from DiaMed AG,
Switzerland in association with Dr. Lapierre, is based on
the Gel Tecnology.

17. Principle of Gel Technology
• Controlled centrifugation of RBCs through a gel column
• Serum and cell reaction takes place in a microtube .
• Six microtubes in a plastic card – easy handling.
• Microtube consists of a reaction chamber that narrows to
become a column with a conical bottom.
• Reaction chamber is designed to allow prior incubation of
test serum and RBCs.
• Each column contains Sephacryl gel suspended in a buffer
solution.
• Depending on the configuration of card , the gel is
premixed with antisera/AHG/other reagents.

18. Add Reactants
Serum/plasma/
red cells
Principle
Reaction
Chamber
Gel and
Reagent
6 Microtubes in Plastic Card

19. Principle of Gel Technology
• Sephadex gel matrix acts as a sieve.
– Large aggutinates remain on or near the top of gel
interface.
– Smaller agglutinates pass partway through gel ,
depending on size.
– Unagglutinated cells pass to base of microtube to
form a button.
• Cells are always added prior to serum so that serum
does not come into contact with gel – this eliminates the
‘WASH PHASE’ as in conventional technique.
• Grading of reaction depending on the distribution of
RBCs throughout the column.

20. Grading of Reaction

21. INTERPRETATION OF GEL TEST
4+4+
Solid band
of red cells
at top of
gel
3+3+
Agglutinated
red cells in
upper half
2+
Red cell
agglutinates
through
length
1+
Aggl. red
cell in
lower half
of gel col.
NEGATIVE

22. Incubator & Centrifuge

23. Uses of Gel Technology
Any immunohaematology test that has
haemagglutination as its end point:
ABO-Rh typing, typing for other blood group systems.
Antibody screening and identification.
Compatibility testing – crossmatching.
DAT/IAT, other Coombs phase test.
Antibody classification- IgG, IgM, IgA, complement…
Specialized hematological tests: PNH, Sickle cell
anaemia.

24. Advantages of Gel Technology
Improved sensitivity and specificity
Easy to use, simple to read
No wash phase in IAT
Minimal training required
Reliable, reproducible results
Easy storage and long shelf life of reagents
Easy disposal of biodegradable cards
Widest range of reagents and instrumentation

25. Disadvantages of Gel
Technology
• Following equipments are required:
1. Special centrifuge to accommodate the microtube
cards
2. Special incubators to incubate the microtube cards
3. Pipette to dispense 25 µl of serum
4. Pipette to dispense 50 µl red cell suspension
• It is expensive
• Skilled technical workers to dispense appropriate
amount of serum or red cell suspension

26. Forward & Reverse typing
• The process of identifying an individual’s bld. grp.
Involves testing of red cells with known antisera
(FORWARD TYPING) and plasma with known group red
cells (BACK/REVERSE TYPING)

27. Forward & Reverse typing
50 µl
Diacell
A1
50 µl
Diacell
B
50 µl
Patient
Serum
50 µl
Patient
Serum
10 – 12.5µl Patients RBC
suspension 5%

28. Glass Bead Technology
• Described in 1993 by Reis K.J.
Principle:
• Test is performed in a microcolumn prefilled with glass
microbeads in suspension of antihuman globulin serum,
any diagnostic reagent or neutral isotonic solution
• Detection of sensitized red cells is based on the seiving
effect of glass microbeads
• Red cells & serum are incubated at the upper part of a
column over the glass microbeads suspension
• These microbeads are calibrated & during centrifugation,
they retain the agglutinates & the unsensitized cells
sediment at the bottom

29. Advantages
• Minimum incubation time of 10 minutes for antibody
screening or cross matching
• Biphasic centrifugation time is only 5 min
• In AHG test there is no need to wash cells or to use
sensitized cells for confirmation
• No tube shaking or resuspension of cell button leading to
variation in reading & grading the agglutination
• Provision of centrifuge calibrated to spin at optimal
speed for fixed & correct length of time reduce error
during this phase
• More objective, consistent & reproducible interpretation
of results

30. Disadvantages
• Special centrifuge to accommodate glass beads
cassettes
• Special incubators to incubate the glass beads cassettes
• Pipettes to dispense 10 µl, 40 µl, 50 µl
• Expensive

31. The ORTHO AUTOVUE INNOVA System is an automated
imunohematology testing system used for blood typing,
antibody screening and compatibility testing using
ORTHOBIOVUE System cassettes. The ORTHO
AUTOVUE INNOVA System is a computer software driven,
fully automated system which provides automated liquid
pipetting,cassette handling, incubation, centrifugation,
reaction grading and interpretation. The software used with
the AUTOVUE INNOVA can be interfaced with a laboratory
information system for test data transfer.

32. DIAGAST-Erythrocytes Magnetised
Technology
Principle:
•Based on magnetization of RBCs in MagneLys magnetizing
solution
•Hemagglutination method is used in combination with a
magnetic field
•Bromelin, a proteolytic enzyme, induces a marked decrease
in the electronegative charge on the surface of RBCs
enabling their agglutination by normally non-agglutinating
antibodies in saline medium
•When subjected to a magnetic field,the magnetized RBCs
migrate & form a pellet at the bottom of the well
•After shaking, free RBCs are resuspended
•Presence of agglutinate shows a positive reaction while
absence of agglutinate shows a negative reaction

33. Advantages
• Quality: Clear-cut reactions to get an unequivocal
interpretation of results
• Comfort: Centrifugation step is completely eliminated
during the analytical process
• Sensitivity comparable to existing techniques

34. Disadvantages
• Stages in the procedure are to be implemented in
succession without interruption
• Only suitably qualified personnel should use the
reagents
• Reactions are to be read at the latest within 2 minutes of
the last shaking

35. TTD Testing
• Compulsory testing for: HIV, HBsAg, HCV, VDRL &
malaria
• Introduction of HBsAg testing in1968
• Introduction of HIV ELISA in1985
• Introduction of HCV testing in 1990
(In India, HCV testing made compulsory from 1997)
• Other TTDs for which screening may be necessary:
CMV, HTLV, prion diseases
• US FDA recommends testing for Anti-HTLV-I & Anti-HBc

36. Various methods for TTD testing:
• ELISA
• Chemiluminiscent microparticle immunoassay (CMIA)
• Nucleic Acid Amplification Testing (NAT)
Automation in TTD testing is available & can be
introduced for all the above methods
Presently at SGH:
HIV, HBsAg, HCV testing: ELISA
Syphilis: VDRL by RPR method
Malaria: Peripheral blood smear

37. Automated CMIA machine for TTD testing

38. Blood Components
Progress in SGH:
• Sassoon Blood Bank was established in 1960
• License for component separation granted in December
1988
• License for preparing saline washed RBCs in 2005
• License has to be renewed every 5 years, Present
License valid upto 2011
• Components available with SGH Blood Bank: Packed
red cells, saline washed RBCs, fresh frozen plasma &
platelet concentrate

39. • Newer techniques in component separation available at
various centres:
- Contact shock freezing: Better & uniform rapid freezing
of plasma
• Apheresis

41. APHERESIS
• Apheresis is collection of anti-coagulated whole blood
from a donor, its separation into components, retention
of desired component and return of remaining
constituents back to the donor with the help of
automated cell separator machines.

42. ADVANTAGES OF APHERESIS
• Reduced multiple donor exposure
– Reduced risk of alloimmunization
– Reduced incidence of transfusion transmitted
diseases
• Full and effective transfusion dose
• Purer product:
– leucocyte reduced products
• High quality product
• Fewer donor reaction due to return of fluid

43. Types of cell separators
 Intermittent flow cell separator (closed system)
 Continuous flow cell separator
 Automated separation techniques by centrifugation
 Cell separation by membrane filtration
 Continuous magnetic cell separator (immunomagnetic)

44. • Automated separation technique by centrifugation:
– Centrifugal force separates blood into different
components depending upon the specific gravity.
– Blood is drawn from an automatic pump
Anticoagulant is added to tube blood is pumped
into rotating bowl, chamber in which layering of
components occurs based on the density desired
component retained and rest returned to donor either
by continuous flow or by intermittent flow.

45. Separation by Membrane Filtration:
– Filtration of plasma through membrane which allows
collection of plasma from a healthy donor.
– Membranes are arranged as hollow fibres which
expels the cellular elements in the flow of blood.
Most commonly used apheresis devices are:
• Haemonetic corporation: Platelets, plasma,
leucocytes.
• Baxter: Plasma, platelets, red cells, leucocyte
• Gambro: Plasma, platelets, leucocyte and
peripheral blood stem cells.

46. TECHNOLOGY FUNCTION
HAEMONETIC Intermittent flow centrifuge separator.
Anticoagulant blood is pumped into rotating bowl
Incoming blood is separated red cells move to the
periphery and plasma to inside of rotating bowl and the
white cells between red cells and plasma
Using optical detectors and fluid surge elutriation process
desired component retained.
GAMBRO(Cobe) •Cobe Spectra: Continuous flow centrifuge cell separator
where two arm technique used- blood is drawn and
returned.
Uses a rotating channel resembling a belt as the initial
separation chamber.
Allows lower WBC and RBC contamination in platelets.
•Cobe TPE: Continuous flow device using a flat
membrane to separate the cells of blood from plasma
BAXTER Continuous flow technology.
CS 3000 has two separation containers firstly for
collection of leucocyte reduced platelets and other for
white cells (CS 3000 plus).

47. HAEMONETICS
CENTRIFUGE

49. COBE Spectra – Automated
Apheresis

50. COBE Spectra: Centrifuge separator

51. BBLOODLOOD SSUBSTITUTESUBSTITUTES
Need
1. To maintain a constant supply of safe alternative
2. To overcome complex procedures of blood collection
and processing
Types
1. Hemopoietic Growth Factors & Pharmacological
Agents
2. Real Blood Substitutes

52. BBLOODLOOD SSUBSTITUTESUBSTITUTES
Hemopoietic Growth Factors
1. They are mainly glycoproteins
2. They are of 3 groups
a. CSF
b. IL
c. Others. Eg. TPO, EPO
3. Can be considered in 4 different functional groups
a. Lineage specific (G-CSF, EPO, TPO)
b. Multi lineage factors (IL3, GM-CSF)
c. Stem cell factors (Steel Factor)
d. Accessory or synergistic factors (IL1, IL6, IL11)

53. RREALEAL BBLOODLOOD SSUBSTITUTESUBSTITUTES
Red Cell Substitutes
A. Hemoglobin Based
Ist
Generation
Purified, unmodified hemoglobin
IInd
Generation
1. Intramolecularly cross linked hemoglobin
2. Recombinant hemoglobin
3. Polymerized hemoglobin (or hemoglobin directly
linked to a high molecular weight non protein
carrier)
IIIrd
Generation
Artificial red blood cell (encapsulated hemoglobin)

54. B. Perflurocarbons
Ist
Generation Fluosol - DA20
IInd
Generation Perfluro-octylbromide
(May improve oxygenation in an infarcted or
ischemic tissue and in tumours)

55. RREALEAL BBLOODLOOD SSUBSTITUTESUBSTITUTES
Platelet Substitutes
Platelet membrane preparation
Freeze dried
Lyophilized platelets
Synthetic platelets
Microspheres of human albumin coated with human
fibrinogen

56. NNEWEW TTECHNOLOGIESECHNOLOGIES IINN BBLOODLOOD
BBANKINGANKING
1. Molecular techniques
Blood group genotyping using allele specific primers
and PCRs for Duffy, Kell, Kidd, MN, ABO, Fetal RhD
typing and HLA typing
2. Gel Technology for immuno-hematology
3. Column Agglutination Technology
4. Erythrocytes Magnetised Technology

57. AAUTOMATIONUTOMATION IINN BBLOODLOOD BBANKINGANKING
• Process whereby an analytical instrument performs many
tests with only minimal involvement of analysts.
– - mechanization of steps in a procedure
• Started in 1940 ; to reduce manual errors due to
fatigue/erroneous sample identification.
• Consolidate chemistry & immunoassay systems on one
platform.

58. • Factors necessitating Laboratory automation
– Turnaround time demands
– Staff shortages
– Less maintenance
– 24 /7 uptime
– Throughput of samples
– ↑ no. of analytes & methods on one system.
– Quality assurance

59. Stages in Laboratory Investigations
• Pre – analytical stage
• Analytical stage
• Post – analytical stage

60. PRE – ANALYTICAL STAGES
IN LAB.
• Sample Delivery -
– Blood drawers or runners / courier facility
– Pneumatic tube delivery system
– Conveyers or track system
– Mobile robots
• Sample processing – Three phases
– Pre-centrifugation :- all measurements in < 45
min.
– Centrifugation :- Blood (Plasma/Serum)
– Post-centrifugation

61. PRE ANALYTICAL MODULES
• Labeling – Bar code label system.
• Sorting – stopper color, size, tests ordered,
instrument design requirements,
• Decapping
• Aliquoting
• Recapping
• Storage / retrieval

62. Automated identification methods
Barcodes
– A barcode comprises a series of vertical bars and
spaces arranged in various combinations to represent
different characters.
– Different barcode systems, each with different rules
governing the representation of the characters. eg.
CODABAR system with ABC symbols
– 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.
– 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

65. SAMPLE IDENTIFICATION

66. CENTRIFUGATION &
VOLUME DETERMINATION

67. DECAPING &
ALIQUOTTING

68. SORTING

69. ANALYTICAL STAGE
• Tasks included –
– Sample introduction & transport to cuvette
or dilution cap.
– Addition of reagent
– Mixing of sample & reagent
– Incubation
– Detection
– Calculation
– Read out & result reporting

70. POST – ANALYTICAL STAGE
• Data processing i.e. – data acquisition, calculation
• Monitoring & displaying – charts & curves
• Performing statistics on patient & control value – flags
• Analyzer computers have capacity to link to –
– Lab. Info. System
– Internet – companies own manufacturing site – can
see real time lab. data & can solve mechanical
problem in short duration.

71. AUTOMATED SYSTEM DESIGNS
• Total Laboratory Automation -
– Japan,1980
– Integration of several instruments = processing
specimen management + transportation systems +
analyzers + digital interpretation + dispatch of results.
• Creates inclusive, continuous network making each
step in testing automated.

72. AUTOMATED SYSTEM DESIGNS
• Disadvantages – Significant financial investment
- Increased floor space
- Highly technical personnel
- Infrastructure remodelling
- Personnel team building
- Software interfacing

73. Blood bank information systems
• Blood bank information systems are computer
systems that have been developed specifically to assist
the blood bank professionals in management of the
patients, donor and blood component information.
• Helps to assess trends and decide future policies by
accessing the statistical information.
• Helps to correlate the laboratory data with donor records
and help to trace the donor records following transfusion
reactions.

74. BBIS consists of :
• Hardware
• Software: application software, operating software and
interface software
• Users
• FDA, AABB and CPA have specific regulatory and
accreditation requirements for BBIS

76. HHAEMOPOIETICAEMOPOIETIC SSTEMTEM CCELLELL TTRANSPLANTRANSPLANT
Types
1. Allogenic
2. Syngenic
3. Autologus
Sources Of Stem Cells
Three main sources:
1. Bone Marrow
2. Peripheral blood
3. Blood in umbilical cord & placenta

77. Indications
 Congenital immune deficiencies
 Severe aplastic anaemias
 Red cell disorders
 Malignant diseases of bone marrow
 Certain solid tumours
 Inherited metabolic disorder
Complications
 Regimen (chemo & radiotherapy) related toxicity
 Rejection
 GVHD: Acute or chronic
 Relapse
 Infectious complications

78. CCORDORD BBLOODLOOD BBANKINGANKING
• Cord blood is defined as blood contained within umbilical
cord & in the placental circulation
• Informed consent preferably from both parents or at least
one parent or a legal guardian
• Collection personnel: Trained obstetrician or trained
nurse, conducting the delivery
Procedure:
• Umbilical cord is clamped & cut as in routine manner
• Insertion site on the umbilical vein is prepared by
cleaning with betadine & alcohol

79. • Sterile needle of the collection bag is inserted into the
umbilical vein pointing towards placenta allowing the
cord blood to flow into the collection bag (with ACD/CPD
anticoagulant) by gravity. Once the vein collapses or the
blood ceases to flow, the collection procedure is
complete.
• On an average, 60-120 ml of anticoagulated cord blood
can be collected.
• Alternatively, heparinised syringe can be used- about 85
ml of blood can be collected & contamination rate is
<1%.

80. • Cryopreservation: In a standard mixture of 20% dimethyl
sulfoxide (DMSO) (10% final concentration), an isotonic
saline or electrolyte solution & a source of protein,usually
autologous plasma or human serum albumin
• Cord blood is vigorously mixed with an equal volume of
the above solution & bags are put in a programmable
freezer at the control rate of -1 to -50
C per minute to -80
to -1000
C & then transferred to liquid nitrogen freezer for
storage at -1960
C
• Stored for a period of 21 years

81. CCORDORD BBLOODLOOD BBANKINGANKING
Advantages
1. Availability
2. Absence of risk to donor
3. Low incidence of viral carriage
4. Recipient tolerance of mismatch
5. Ethnic targeting possible
Disadvantages
• Ethics and consent issues
• Cost varying from Rs.50,000 to Rs.1 lakh depending
on the private stem cell bank involved
• Lack of implementation of proper guidelines in cord
blood banking

82. Bone Marrow
Transplantation
Peripheral blood stem cell
transplantation
Procedure painful, carried
out under spinal or general
anaesthesia
Procedure less painful,
obtained by apheresis & can
be done on an outdoor basis
Need to filter out most of
the bone chips, clots &
fibrin
PBSCs are cleaner, bone &
tissue debris from bone
marrow harvesting are
eliminated
Recovery comparatively
slower
Patients recover more quickly

83. References
• WHO Transfusion Medicine Technical Manual, 2nd
Edition 2003
• Internet:
- NACO Blood Bank (PDF Document)
- FDA Guidelines to Inspection of Blood Banks
- Voluntary Blood Donation Programme: Operational
Guide by NACO
• DiaMed & DiaGast Technical Manual

84. THANK
YOU