1. • IMMUNOLOGY - study of the immune system
• SEROLOGY- testing of antigen and or antibody levels of the
blood
Specimen Preparation & Collection
o Most frequently encountered specimen in
immunological testing is: SERUM
o Fresh, nonheat inactivated serum
o May be stored between 2°C and 8°C for upto 72 hours.
o If there are any delays in testing, the serum should be
frozen at -20°C or below.
o Blood is collected antiseptically by venipuncture.
o Avoid hemolysis- may cause false positive results.
o Blood specimen is allowed to clot at RT or at 4°C and
then centrifuged.
o Serum should be separated into another tube without
transferring any cellular elements.
Antigens, Immunogens, and Antibodies
o An ANTIGEN is considered as any foreign particle that enters
the body such as dust, food and any microorganisms.
o An IMMUNOGEN is any foreign particle that triggers a
response from the body's immune system.
o An ANTIBODY is a blood protein produced in response to
and counteracting a specific antigen. Antibodies combine
chemically with substances that the body recognizes as
alien, such as bacteria, viruses, and foreign substances in
the blood.
o 5 classes of antibody
– IgG, IgA, IgM, IgD, IgE
GENERAL FUNCTIONS:
1. Neutralize toxic substances.
2. Facilitate phagocytosis.
3. Combine with antigens on cellular surfaces
and thereby cause destruction of these cells
either intravascularly or extravascularly.
AGGLUTINATION
o A reaction in which particles (as red blood cells or
bacteria) suspended in a liquid collect into clumps and
which occurs especially as a serological response to a
specific antibody.
o Lattice formation
LATERAL FLOW TESTS
o are simple devices intended to detect the presence (or
absence) of a target analyte in sample (matrix) without
the need for specialized and costly equipment, though
many lab based applications exist that are supported
by reading equipment.
1 MT 35: Laboratory Management in Immunology-Serology & BloodBanking by: Filconey Rose B. Camado, RMT
2. IMMUNOHEMATOLOGY
o more commonly known as blood banking
o branch of hematology which studies antigen-antibody
reactions and analogous phenomena as they relate to
the pathogenesis and clinical manifestations of blood
disorders.
Blood Bank Station
o A place where blood is collected from donors, typed,
separated into components, stored, and prepared for
transfusion to recipients.
o may be a separate free-standing facility or part of a
larger laboratory in a hospital.
Chromosomal Locations of Major Blood Group Systems
CHROMOSOME LOCUS
1 Rh, Duffy
4 MNS
7 Kell
9 ABO
18 Kidd
19 H, Lewis, Lutheran
22 P1
ABO Blood Group System
o Karl Landsteiner (1901)
o discovered that different people's blood had different
characteristics that made it "incompatible" with other
people's blood that didn't carry those same traits.
o discovered the A, B, and O blood types.
o discovery of the differences and identification of the
groups that were alike made it possible for blood
transfusions to become a routine procedure.
o paved the way for many other medical procedures
that we don't even think twice about today, such as
surgery, blood banks, and transplants.
ABO Forward Grouping
BLOOD GROUP REACTION W/
ANTI-A
REACTION W/
ANTI-B
A + -
B - +
AB + +
O - -
ABO Reverse Grouping
BLOOD GROUP REACTION W/ A
Cells
REACTION W/ B
Cells
A - +
B + -
AB - -
O + +
Red Cell Antigen-Antibody Reactions
SEROLOGIC GRADING – MACROSCOPIC EVALUATION
4+ One solid agglutinate
3+ Several large agglutinates, clear background
2+ Medium-sized agglutinates, clear background
1+ Small agglutinates, turbid background
W+ Tiny agglutinates, turbid background
2 MT 35: Laboratory Management in Immunology-Serology & BloodBanking by: Filconey Rose B. Camado, RMT
3. 0+
No agglutination or hemolysis
Agglutination in the Gel Test
4+ Agglutinated cells form a cell
layer at the top of the gel
media.
3+ Agglutinated cells begin to
disperse into the gel media
and are concentrated near
the top of the microtube.
2+ Agglutinated cells disperse
into the gel media and are
observed throughout the
length of the mirotube.
1+ Agglutinated cells disperse
throughout the gel media and
may concentrated toward the
bottom of the microtube.
Negative All cells pass through the gel
media and form a cell button
at the bottom of the
microtube.
Mixed-field Agglutinated cells form a
layer at the top of the gel
media. Unagglutinated cells
pass to the bottom of the
microtube.
Donor Selection
1. Appears to be in good health.
2. Age: 18-65 years old
3. Body weight : Max of 10.5 mL bld/kg
Ideal : 10lbs. or 50kgs can donate
a. 450 mL bld + 30 mL bld (serotest)
4. Temperature
5. Pulse
6. Blood pressure
7. Minimum hemoglobin and hematocrit
Donation Process
1. Donor Registration
2. Interview and Physical Examination
3. Donor selection and Blood collection
a. Donor Bleeding
• 7-10 minutes/ <15minutes
• Angle : 45º angle reduce to 10º-20º once in
skin
• Blood Pressure Cuff used as torniquet:
o Set at 40-60 mmHg
Donor Deferral
A. Permanent
Chagas, Babesiosis
Tegison for Psoriasis (teratogenic)
Risk of CJD
Recepient of pituitary derived Growth
Hormone
a. Human derived GH – permanent
b. Recombinant GH – no deferral
Recipient of cornea and Dura mater transplant
B. 3-year deferral
3 MT 35: Laboratory Management in Immunology-Serology & BloodBanking by: Filconey Rose B. Camado, RMT
4. Malaria
• Refugee or immigrant of an endemic area of
Malaria
C. 1-year or 12 month deferral
Tattoo
Rabies vaccine
Traveller of an endemic place with Malaria
Blood transfusion
Major operation including Dental Surgery
a. Dental Surgery – 1 year
b. Tooth Extraction – 3 days
D. Recent Blood Donation
a. AABB – 8 weeks/ 2 months
b. PHILIPPINES – 12 weeks/ 3 months
450 mL blood – 12 weeks
200 mL blood – 6-8 weeks
E. Childbirth
a. AABB – 6 weeks
b. PHILIPPINES – 9 months after childbirth
F. 4-week/1 month deferral
o German measles vaccination (Rubella)
o Tx w/ isotretinoin (Accutane) for Acne
o Proscar for BP
G. 2-week deferral
o Measles vaccination (Rubeola)
o Typhoid vaccination
H. 2-day deferral
o After hemapheresis
I. 2-3-week deferral
o After febrile episode
J. 12-24-hour deferral
o After recent alcohol intake
K. Typhoid
a. Infection – 3-month deferral
b. Household contact – 4 weeks from last
exposure
Blood Bag
1 part anticoagulant : 7 parts blood
a. 63 mL AC – 450 mL bld
b. 31.5 mL AC – 200 mL bld
Blood Preservation: Anticoagulants and Red Cell Additives
Citrate - binds Calcium
Dextrose - provide energy for red cell
Citric acid - prevents caramelization
Phosphate buffer – inc. ATP levels
Adenine - for improved survival of RBCs
Approved Preservatives
Acid-citrate-dextrose (ACD) 21 days
Citrate-phosphate-dextrose
(CPD)
21 days
Citrate-phosphate-adenine
(CPDA-1)
35 days
Citrate-phosphate-double 21 days
4 MT 35: Laboratory Management in Immunology-Serology & BloodBanking by: Filconey Rose B. Camado, RMT
