LAB # 3 - INDICATOR: RBC (Red blood cell)

- Classic pregnancy agglutination reaction

AGGLUTINATION - DETECT: Human Chorionic Gonadotropin
- SAMPLE: Urine
REACTION 5. Coaagglutination
Agglutination - uses bacteria as the inert particles to which
 Agglutination is the visible aggregation of antibody is attached
particles caused by combination with - Staphylococcus areus is most frequently
specific antibody. used, because it has a protein on its outer
 Naturally attached & Synthetically attached surface, called Protein A  absorb FC
 Antibodies that produce such reactions are region of antibody leaving FAB region
often called “agglutinins”. available on environment to react with
 Agglutination involves a two-step process: specific antigen
 Sensitization – unite through
6. AHG-Mediated agglutination: AKA “Coombs
antigenic determinants
Test”
 Lattice formation – rearrangement of
- detects nonagglutinating antibody by means
antigen-antibody bonds to form
of coupling with a second antibody  IgG
stable lattice.
with Anti-Human Globulin Reagent)
 Types of particles participating in such
- The Coombs’ test can be divided into two
reactions include erythrocytes, bacterial
different types, direct and indirect, each of
cells, and inert carriers such as latex particles
which has a different purpose
(common).
 IgG & IgM (more effective – **Positive reaction
structure) Direct Coombs test/ Indirect Coobs test/IAT
 Agglutination reactions can be classified into DAT
several distinct categories:  HDN – Hemolytic  Cross-matching
1. Direct agglutination Disease of Newborn  Antibody detection
- occurs when antigens are found naturally on  Antibody
a particle  HTR – Hemolytic identification
- If an agglutination reaction involves red Transfusion Reaction  RBC Antigen
blood cells, it is called Hemagglutination  AIHA – Auto- phenotyping
2. Passive agglutination immune Hemolytic
- Employs particles that are coated with Anemia
antigens not normally found on their surfaces
- DETECT: Antibody
- SAMPLE: Serum
- Antigen is accompanied by a carrier particle
 Latex, charcoal
- ASO (Anti-Streptolysin O), RF (Rheumatoid
Factor), ANA (SLE) (Anti-nuclear Antigen)
3. Reverse passive agglutination
- Employs particles that are coated with
antibodies
- DETECT: Antigen
- Antibody is accompanied by a carrier
particle
- CRP latex agglutination
Carrier Particle
4. Agglutination inhibition  For antibody
- based on competition between particulate  Should have the ability to absorb FC region
and soluble antigens for limited antibody- and not FAB
combining sites, and a lack of agglutination  Latex, charcoal
is an indicator of a positive reaction
DIRECT AGGLUTINATION
  EXAMPLE→ ABO forward typing
 Detects the presence of ABO antigen using
ABO anti-sera (Anti-A -blue and Anti-B
-yellow)
 Slide and tube method
Pre-requisites:
 Principle of Agglutination reaction
 Centrifugation
 REAGENTS: Anti-A and Anti-B
(Monoclonal)
Slide Method: Procedure
1. Label the glass slides as Anti-A and Anti-B
2. Add 1 drop of whole blood from a capillary
puncture to each drop of the typing antiserum.
NOTE: Samples from EDTA tube can also be
used.
Shake 6-8 times.
3. Place 1 drop of anti-A and 1 drop of anti-B
reagent separately on a labeled slide.
NOTE: AVOID CONTAMINATING THE
TIP OF THE DROPPER OF THE
REAGENT WITH PATIENTS SAMPLE.
4. Mix the cells and reagent using a clean
applicator stick. Spread each mixture evenly
(circular motion) on the slide over an area of
10-15 mm diameter.
5. Tilt the slide for 2 minutes at room
temperature (22◦C - 24◦C) and observe for
agglutination. NOTE: Do not read results
after 2 minutes.
6. Read and Record the result.
 Positive reaction➔ With
Agglutination
 Negative Reaction➔ Without
Agglutination

**NOT ALWAYS RBC, it can also be:

- Bacterial Suspension
 Wethal
 Weil Felix