Agglutination Flashcards
• It is actually a foreign substance
Antigen
• Substance that is produced against the antigen
Antibody
• particulate antigens are linked together with its corresponding
antibodies
Agglutination
• Used as indicator cells
Red cell suspension
• Demonstrates Ag-Ab reactions
Red cell suspension
soluble which is difficult to see
Ag-Ab reaction
Specific specimen of the RCS
Anti-coagulated blood
(purple=EDTA=anticoagulant)
Packed RBC(after centrifugation)
Preparation of Red Cell Suspension
Wash 3x for 5 mins 1500 rpm
What does washing remove
Cytokines
Unwanted antibodies that may interfere with antigen-antibody reaction.
Micro-aggregates,
Plasma
What will the antibodies that are still present cause?
Premature reaction rxn
False negative rxn
- An isotonic solution
- Preserves the morphology of RBCs
- Preferred because of economical advantage
NSS (0.9% Normal Saline Solution)
Other suspending agents
PBS (Phosphate Buffer Saline)
Ideal RCS
• 2 to 5 % Red Cell Suspension
Vital
Thin suspension
For microtiter plates
reduce the concentration of RCS
not a good indicator cell
Thick suspension
First step:
Add 2 drops of whole blood to the wasserman
Second step
Wash the Cells 3x with 0.85% NSS
Third step
Add NSS until the suspension will
become CHERRY RED color
RCS should always be made fresh (using freshly extracted blood)
RCS should always be made fresh (using freshly extracted blood)
Shelf life of RCS
Used within 24 hours
Preservatives to be used:
Glycerol
Process of progressively reducing or lowering the concentration of
solute in a given solution
Serial Dilution
Measures the antibody concentrations in test serum
Serial Dilution
High titer
high infection/high immunity
Varying antigen
Volume of antibody is constant
Alpha
procedure (Dean Webb procedure)
Varying antibody
Volume of antigen is constant
Beta procedure (Ramon procedure)
Excess of antibody/extremely high titer reacting to small amount of antigen
HIGH ANTIBODY
Prozone reaction
Excess of antigen/low titer reacting to a very small amount of antibody
LOW antibody
Post zone reaction
Primary Phenomenon
Sensitization
Binding of Ab to Ag
Secondary Phenomenon
Lattice formation
2 Fab
Tertiary Phenomenon
Detected by effect on tissues or cells
DF
TV/SOLUTE