Blood banks and Blood

grouping
BLOOD BANKS
• Blood banks collect, test, and store blood.
• Autologous transfusion - If surgery is scheduled months in advance,
patients may be able to donate their own blood and have it stored.

blood and blood transfusions 2

SELECTION CRITERIA OF BLOOD
DONORS
• The criteria for donor acceptance are:
(a) Age : between 18 to 65 years.
(b) Gender: Male and Female.
(c) Weight : Minimum 45 kg
(d) Donors must not have medical history that could harm
themselves or the potential recipient of their blood. These
include inherited bleeding disorders, recent illness or
consumption of medication.
(e) Haemoglobin level: 12.5g/dl – 18.0g/dl.

blood and blood transfusions 3

(f) The interval between the last donations of whole blood should
not be less than 8 weeks and not less than 2 weeks for plasma or
platelet donation.

(g) Temperature and pulse of the donor should be normal.

(h) The systolic and diastolic blood pressures are within normal limits
without any medication.

(i) The arms and forearms of the donor should be free from skin
punctures or scars indicative of professional blood donor or
addiction of self injected narcotics.
4
Blood Collection
1. Donor Identification:
- A careful check must be made to verify the Donor’s identity against
the Blood Donor Enrolment Form.
- Verify the name, blood group and barcode number on the Blood
Donor Enrolment Form with the labels on the blood bags at the
bedside before venepuncture.

5
2. Need for successful venepuncture and proper mixing of blood.
- Needle should be inserted into the vein at first attempt.
- Proper mixing of the blood with the anticoagulant should be
done.
- Blood must be immediately mixed: Flow of the blood must be
sufficient and uninterrupted.
- Manual mixing of blood bags must be inverted every 30 – 45
seconds.
- When an automated blood mixer is used, an appropriate
validation system is required.

6
blood and blood transfusions 7
3. Bleeding should NOT be longer than 12 minutes.
- If the bleeding time is longer than 12 minutes it should not be
used for platelet preparation.

4. At the end of donation take blood samples directly from the

donor venepuncture tubing into the sample tubes. DO NOT SQUEEZE
FROM THE
BLOOD BAG.

blood and blood transfusions 8

Adverse reactions in Donors
• Giddiness/ syncope (vasovagal syndrome)
• Convulsions
• Vomiting
• Tetany/ muscle spasm
• Hematoma
• Eczematous reactions of the skin around venupuncture.
• Delayed syncope

blood and blood transfusions 9

Handling of filled containers and samples
• Containers should be checked before and after donation for any
defect.
• Verification of blood sample container with donor’s identification.
• Labelling and withdrawal of blood samples at bedside.

blood and blood transfusions 10

 Labeling of sample

• Write clearly:
- unit no.
- date of collection and expiry
- Volume

• After the bags are labelled ask a second technician to double check the number
and group on the bags.

• Label those found unsuitable for use with a biohazard label and keep for disposal.
-

11
BLOOD STORAGE
• Blood products must be stored at 4C +- 2C.
• Stored blood has a shelf life of 3 weeks.
• After a storage time of 24-72 hr RBCs have reduced capability to
release oxygen to tissues.
• If the patient needs massive transfusions its better to give blood that’s
less than 7 days old.

12
 Issue of blood and blood products
• Blood may be released by the blood bank on request or may be
stored in a designated blood refrigerator to be released by the blood
bank staff to an authorized staff.

• Blood Bank staff must ensure that the correct blood and blood
product is issued out to the requesting person.

13
 BLOOD COMPONENTS

Whole Blood

Slow Centrifugation

Packed Red Blood Cells Platelet Rich Plasma

High Speed Centrifugation

1 Unit of Random Donor 1 Unit of Fresh Frozen Plasma

Platelets
Thawing precipitates the
plasma proteins

Cryoprecipitate
Warming Blood

• STORED BLOOD IS COLD 4*C

• PATIENTS UNDERGOING SURGERY WILL ALREADY BE
LOSING BODY HEAT DUE TO WOUND OR CAVITY
EXPOSURE
• LARGE VOLUMES OF COLD BLOOD MAY INDUCE
HYPOTHERMIA OR CARDIAC ARYTHMIA

blood and blood transfusions 15

• WARMED BLOOD IS MOST COMMONLY REQUIRED IN LARGE VOLUME RAPID TRANSFUSIONS &
EXCHANGE TRANSFUSION IN INFANTS. (>50 ml/kg/hr in adult and >15ml/kg/hr in children)

• BLOOD SHOULD ONLY BE WARMED IN A BLOOD WARMER THAT HAVE A VISIBLE THERMOMETER AND
AN AUDIBLE WARNING ALARM AND SHOULD BE PROPERLY MAINTAINED.

• Blood must NOT be warmed by placing it into hot water, in microwave, on radiator, under running
water or near any uncontrolled heat source.

• Blood which has been warmed must not be re-refrigerated for later use or reissued.
• Warming tends to accelerate red cell metabolism producing haemolysis and may permit bacterial
growth.

blood and blood transfusions 16

Blood Transfusion Administration

blood and blood transfusions 17

Pre-administration Procedure
• Step 1: Check the blood component
has been prescribed
• Step 2: Undertake baseline
observations
• Step 3: Undertake visual inspection
LEAKS
DISCOLOURATION
CLUMPING

EXPIRY DATE

blood and blood transfusions 18

• Remain with the patient during
the first 15-30 minutes of the
infusion.
• Infuse the blood product at the
prescribed rate.
• Monitor vital signs.

blood and blood transfusions 19

 Types of BT
• Based on time of transfusion
• Fresh whole blood transfusion
• Stored CPD Blood

• Based on composition
• Whole blood
• Blood fraction

• Based on the donor

• Autologous blood transfusion
• Blood from diff donor

20
Whole Blood
• Storage
• 4° for up to 35 days
• Indications
• Massive Blood Loss/Trauma/Exchange Transfusion
• Considerations
• Donor and recipient must be ABO identical
RBC Concentrate
• Storage
• 4° for up to 42 days, can be frozen
• Indications
• Many indications—ie anemia, hypoxia, etc.
• Considerations
• Recipient must not have antibodies to donor RBC’s (note: patients can
develop antibodies over time)
• Usual dose 10 cc/kg (will increase Hgb by 2.5 gm/dl)
• Usually transfuse over 2-4 hours (slower for chronic anemia)
Platelets
• Storage
• Up to 5 days at 20-24°
• Indications
• Thrombocytopenia, Plt <15,000
• Bleeding and Plt <50,000
• Invasive procedure and Plt <50,000
• Considerations
• Contain Leukocytes and cytokines
• 1 unit/10 kg of body weight increases Plt count by 50,000
• Donor and Recipient must be ABO identical
Plasma and FFP
• Contents—Coagulation Factors (1 unit/ml)
• Storage
• Comes in 200ml bags.
• FFP--12 months at –18 degrees or colder
• Indications
• Coagulation Factor deficiency, fibrinogen replacement, DIC, liver disease, exchange transfusion,
massive transfusion
• Considerations
• Plasma should be recipient RBC ABO compatible
• In children, should also be Rh compatible
• Account for time to thaw
• Usual dose is 20 cc/kg to raise coagulation factors approx 20%
 Cryoprecipitate

• Description
• Precipitate formed/collected when FFP is thawed at 4°
• Storage
• After collection, refrozen and stored up to 1 year at -18°
• Indication
• Fibrinogen deficiency or dysfibrinogenemia
• vonWillebrands Disease
• Factor VIII or XIII deficiency
• DIC (not used alone)
• Considerations
• ABO compatible preferred (but not limiting)
• Usual dose is 1 unit/5-10 kg of recipient body weight
Granulocyte Transfusions
• Prepared at the time for immediate transfusion (no storage available)
• Indications – severe neutropenia assoc with infection that has failed
antibiotic therapy, and recovery of BM is expected
• Donor is given G-CSF and steroids
• Complications
• Severe allergic reactions
• Can irradiate granulocytes for GVHD prevention
Leukocyte Reduction Filters
• Used for prevention of transfusion reactions
• Filter used with RBC’s, Platelets, FFP, Cryoprecipitate
• Other plasma proteins (albumin, colloid expanders, factors, etc.) do
not need filters—NEVER use filters with stem cell/bone marrow
infusions
• May reduce RBC’s by 5-10%
• Does not prevent Graft Verses Host Disease (GVHD)
RBC Transfusions
Preparations
• Type
• Typing of RBC’s for ABO and Rh are determined for both donor and recipient
• Screen
• Screen RBC’s for atypical antibodies
• Approx 1-2% of patients have antibodies
• Crossmatch
• Donor cells and recipient serum are mixed and evaluated for agglutination
Platelet Transfusions
Preparations
• ABO antigens are present on platelets
• ABO compatible platelets are ideal
• This is not limiting if Platelets indicated and
type specific not available
• Rh antigens are not present on platelets
• Note: a few RBC’s in Platelet unit may
sensitize the Rh- patient
 Platelet Transfusions
Administration
• Dose
• May be given as single units or as apheresis units
• Usual dose is approx 4 units/m2—in children using 1-2
apheresis units is ideal
• 1 apheresis unit contains 6-8 Plt units (packs) from a single
donor
• Procedure
• Should be administered over 20-40 minutes
• Filter use
• Premedicate if hx of Transfusion Reaction
• Complications—Transfusion Reaction
Autologous Blood Transfusions
• Collection/infusion of client’s own blood
Four types:
• Preoperative autologous blood donation
• Acute normovolemic hemodilution
• Intra-operative autologous transfusion
• Postoperative blood salvage

blood and blood transfusions 31

Preoperative autologous blood donation
• Collecting whole blood from the client, dividing it into components
and storing it for later use
• Can be collected weekly.
• Can be stored up to 40 days; up to 10 years for rare blood types.

blood and blood transfusions 32

Acute normovolemic hemodilution
• Withdrawal of client’s RBCs and volume replacement just before a
procedure
• Goal is to decrease RBC loss during surgery
• Blood is stored at room temperature for up to 6hrs and reinfused
after surgery.
• Not for anemic clients or those with poor kidney function.

blood and blood transfusions 33

Intra-operative autologous transfusion & Post
operative blood salvage
• Recovery/reinfusion of client’s own blood from operative field
or bleeding wound.
• Special devices collect, filter, drain blood into transfusion bag
• Used for trauma or surgical patients with severe blood loss
• Blood must be reinfused within 6 hours.

blood and blood transfusions 34

Blood grouping
 Human Blood Groups

• Discovered by Landsteiner in 1901

• Blood group antigens are structures on the outer surface of

human red blood cells (RBCs) that can be recognized by the
immune system of individuals who lack that particular
structure.

• Presence or absence of these antigens is used to classify

blood groups.
Human Blood Groups

• The International Society of Blood Transfusion recognizes

35 blood group systems and 6 antigen collections

• Each blood group system consists of a group of antigens

encoded by alleles at a single gene locus or very closely
linked loci.

Source : Williams Hematology Ninth Edition,

Copyright © 2016,
Human Blood Groups

• Major blood groups – ABO & Rh

• Minor blood groups –

 MNS system
P
 Lutheran
 Kell
 Lewis
 Duffy
 Kidd
 Diego
 Hh/Bombay
 Kx ……..
Human Blood Groups
Human Blood Groups

The main clinical importance of a blood group system:

• depends on the capacity of alloantibodies (directed against

the antigens not possessed by the individual) to cause
destruction of transfused red cells.

• To cross the placenta and give rise to haemolytic disease in

the fetus or newborn.

• On these criteria, the ABO and Rh systems are of major

clinical importance.
ABO Blood Group

• Discovery of the ABO system by Landsteiner marked the

beginning of safe blood transfusion.

• Discovered by Karl Landsteiner in 1901

• When he noticed that the red cells of some individuals

could be agglutinated by the serum of others.
ABO Blood Group
Landsteiner’s Rule

• If an antigen (Ag) is present on a patients red blood cells the

corresponding antibody (Ab) will NOT be present in the
patients plasma, under ‘normal conditions’.

ABO blood group consist of

• two antigens (A & B) on the surface of the RBCs
• two antibodies in the plasma (anti-A & anti-B)

BLOOD GROUP =B
ABO Blood Group

• There are four main blood groups: A, B, AB and O

• In Caucasian population, the frequency of group A is 42%,

B 9%, AB 3% and O 46%.
ABO Blood Group
 ABO Blood Group
“Genetics”

Three alleles of the ABO gene (A, B, and O)

A and B are co-dominant alleles.

O is recessive allele.

Four different phenotypes: A, B, AB and O

Six possible genotypes: AA, AO, BB, BO, AB and OO
Formation of the H antigen

RBC

Glucose

H antigen Galactose

N-acetylglucosamine

Fucose Galactose
 ABO BLOOD GROUP
“GENETICS”
• The H antigen is the foundation upon which A and B
antigens are built.

• A and B genes code for enzymes that add an

immunodominant sugar to the H antigen.

• Immunodominant sugars are present at the terminal ends

of the chains and confer the ABO antigen specificity.
 ABO BLOOD GROUP

• The “A” gene codes for an enzyme (transferase) that

adds N-acetylgalactosamine to the terminal sugar of the
H antigen

N-acetylgalactosaminyltransferase

• The “B” gene codes for an enzyme that adds D-

galactose to the terminal sugar of the H antigen
D-galactosyltransferase
Formation of the A antigen

RBC

H Antigen
Glucose

Galactose

N-acetylglucosamine

Fucose Galactose

N-acetylgalactosamine
Formation of the B antigen

RBC

H Antigen
Glucose

Galactose

N-acetylglucosamine

Fucose Galactose

D-Galactose
 ABO Blood Group

• The O gene does not encode a functional enzyme;

• Hence, a specific sugar is not attached to the red cell

membrane .
 Bombay Phenotype (Oh)

• The individual is homozygous for the inactive h allele of FUT1 and

hence cannot form the H precursor of the A and B antigen.

• Their red cells type as group O, but their plasma contains anti-H in
addition to anti-A, anti-B and anti-A,B, which are all active at 37 °C.

• As a consequence, individuals with an Oh Bombay phenotype can

only be safely transfused with other Oh red cells.

• Serologically, the red cells group as O; unlike the true O phenotype,

red cells from Bombay phenotype lack H antigen
ABO Blood Group

Blood Group Antigen H Substance Antibody

A A antigen + Anti-B

B B antigen + Anti-A

AB AB antigen + No Antibody

O H antigen ++ Anti-A anti-B

Bombay blood group None No H substance Anti-H anti-A

Anti- B
 Rh system

The Rh system, formerly known as the Rhesus system, was

so named because the original antibody that was raised by
injecting red cells of rhesus monkeys into rabbits and
guinea pigs reacted with most human red cells.

• Discovered by, Landsteiner and Wiener In 1940.

 Rh group

• Rh is a blood group system with many antigens, one of

which is D (RH1) .
• Presence or absence of blood antigen D

Unlike the ABO system, individuals who lack the D antigen

do not naturally produce anti-D.

Production of antibody to D requires exposure to the

antigen.
 Clinical Significance of Rh blood
groups

Hemolytic disease of newborn

• when there is incompatibility of blood group between
mother and baby

• Known as erythroblastosis foetalis.

• occurs when mother is Rh –ve ( dd) and father is Rh+ve

resulting in fetus which is Rh +
Hemolytic disease of newborn
 Hemolytic disease of newborn

• The chance of the first pregnancy to suffer from hemolytic

disease of newborn is relatively small

• In subsequent pregnancies: the risk of developing Ab (Anti –D

IgG) increases causing hemolytic disease of newborn

• So anti-D Ig prophylaxis is given.

-Anti–D Ig (Rhogam) administered to Rh-ve mothers at 28

weeks and within 72 hrs of delivery.
Also administered following abortions
 Serologic Detection of Erythrocyte Antigens and
Antibodies
2 components :

• Test unknown cells with known antibodies (forward)

• Test unknown serum/plasma with known red cells
(reverse)

• The patterns are compared and the blood group is

determined.

• Positive reactions are seen as hemagglutination or

hemolysis, and the results of one test should confirm the
results of the other.
 Agglutination of red cells by antibody

• Carried out in tubes, microtitre plates or using column

agglutination (gel) technology, centrifugation or
sedimentation.

• Slide tests are used for emergency ABO and D grouping

 Blood sample

• Clearly labeled blood samples in sterile tubes (plain & EDTA).

• Test should be performed on the fresh sample for best results.

• If serum is not completely separated, centrifuge tube at 1000-3000

rpm for 3 min.

• No signs of hemolysis should be there

• Preferably use saline washed red cells and make 2-5% red cell
suspension.
Anti- serum
 Slide tests.

• These are used rarely in a few parts of the world.

• Because of evaporation, slide tests must be read within about 5

min.

• Reagents that produce strong agglutination within 1–2 min are

normally used for rapid ABO and RhD grouping.

• Because the results are read macroscopically, strong cell

suspensions should be used (50% cells in their own serum or
plasma).
 Slide Method for ABO grouping

• Take a slide and label A, B and D.

• Put 1/1 drop of 40-50%
suspension of test red cells on
labeled slide
• Add 1/1 drop of anti-A, anti- B and
anti-D to respective drop of blood
as labeling.
• Mix the contents.
• Tilt the slide gently back and forth.
• Examine for agglutination
 Slide tests.

ADVANTAGES:
• – Preliminary typing tests

DISADVANTAGES:
• – Not routine test
• – Less sensitive
• – Drying of reaction giving to false positive results
Slide tests.
Anti-A Anti-B Anti-D
 Test Tube Method of ABO Grouping
Recommended method

• Allows longer incubation of antigen and antibody mixture without drying

• Tubes can be centrifuged to enhance reaction
• Can detect weaker antigen / antibody

Two steps in ABO grouping

• Cell grouping (Forward grouping)

Tests the patients red cells with known Anti-A & Anti-B to determine the antigen
expressed

• Serum grouping (Reverse grouping)

Test the patients serum with known A & B cells to determine the presence of
antibody
Lay Out of Tubes for ABO & Rh grouping
 2 vol of anti- A/ 1 vol of 2-5%
anti-B/ red cell
anti-D suspension
Incubate at room temp
(20-24oC) for 5 min

Forward Centrifuge at 1000 rpm

for 1 min
Grouping

Check for agglutination

against well lighted
background O+ve
 1 vol of 5%
2 vol of test
suspension of
serum/
reagent red cells
plasma
in respective
tubes

Shake & leave at room

temp (20-24oC) for 5 min

Reverse
Grouping Centrifuge at 1000 rpm
for 1 min

Centrifuge & record the

results similarly as for
“O”
cell grouping
Recording results of ABO grouping

Reaction of red cells Reaction of serum Interpretation

with with pooled cells
Anti-A Anti- Anti-D A B O
B cell cell cell
+ 0 + 0 +/H 0 A +ve
+ 0 0 0 +/H 0 A-ve
0 + + +/H 0 0 B+ve
0 + 0 +/H 0 0 B-ve

+ = agglutination, 0 = no agglutination H = hemolysis

Recording results of ABO grouping

Reaction of red cells Reaction of Interpretation

with serum with
pooled cells
Anti-A Anti- Anti-D Ac Bc Oc
B
+ + + 0 0 0 AB+ve
+ + 0 0 0 0 AB-ve
0 0 + +/H +/H 0 O+ve
0 0 - +/H +/H 0 O-ve
0 0 +/H +/H + Oh

+ = agglutination, 0 = no agglutination H = hemolysis

Grading of Agglutination
Grading of Agglutination
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