TYPES OF AGGLUTINATION REACTION

DIRECT AGGLUTINATION

PASSIVE AGGLUTINATION

REVERSE PASSIVE AGGLUTINATION

AGGLUTINATION INHIBITION

COAGGLUTINATION

AHG-MEDIATED AGGLUTINATION

6 types of agglutination reaction

DIRECT AGGLUTINATION

Occurs when antigens are found naturally on a particle.

KNOWN BACTERIAL ANTIGENS

________ ___________ _________ - used to test for the presence of unknown antibodies in the patient.

HEMAGGLUTINATION

IgM

If an agglutination reaction involves red blood cells, it is called (a)___________________.

(b) ____ is the ab responsible for this reaction.

(a) _______________ is routinely done at room temperature withoutenhancement techniques.

ABO BLOOD TYPING

___ _______ _______ - clumping on the surface area of the RBC.

ANTI-A

ANTI-B

ROOM TEMP

Anti-A and anti-B antisera are IgM in nature, that’s why the best temperature to incorporate when performing the ABO blood typing is at ____ ______.

IgM

___ - cold reacting antibody. Best reacts at room temp or colder.

SLIDE METHOD

TUBE METHOD

Two methods: (a) _______ __________ and (b)_____ __________.

Gold standard for forward typing is the (b) _____ ________.

GLASS SLIDES

TEST TUBES

Direct agglutination can be done on (a)______ _______ or (b)_______ _______.

• (b)_______ _______ also can be centrifuged and then sh-ken to see if the cell can be evenly resuspended.

• The degree of agglutination is the graded.

• The tubes should be tilted qhen grading for better visualization.

PARTIAL

COMPLETE HEMOLYSIS

In serology, (a)________ or (b) _________ __________ is considered also a positive reaction.

4+

3+

2+

1+

NEGATIVE

• __ - one solid clump

• __ - severaral large clumps

• __ - intended for numerous smaller clots

• __ - barely discernible clumps

• ________ – smooth suspension

HEMOLYSIS

NO HEMOLYSIS

COMPLETE HEMOLYSIS

PARTIAL HEMOLYSIS

Need to compare to (a)___________. (a)____________ is considered as a positive reaction.

• (b) ___ ____________ – intact cell button with clear supernatant.

• (c) ___________ ____________– absent cell button with red supernatant.

• (d) ___________ ____________ – present cell button, supernatant is pink in color

WIDAL TEST

SOMATIC ANTIGEN (O)

FLAGELLAR ANTIGEN (H)

(a)_______ _______ - a rapid screening test to help determine the possibilityof salmonellosis or typhoid fever

• The antigens used in this procedure include Salmonella (b)__________ _________ ___ and the (c)___________ _________ ___ of S.typhi serovar typhi.

WEIL-FELIX TEST

PROTEUS SPP.

(a)_____-______ _____ - intended for Rickettsial disease

• (b)_________ ____ – same antigenic determinants to Rickettsial diseases (cross reaction mechanism)

• OX-2, OX-19 OX-K

PASSIVE AGGLUTINATION

• Also known as Indirect Agglutination

• Employs particles that are coated with antigens not normally found on their surfaces

ANTIBODY

ANTIGENS

Detects: (a) __________

Reagant: (b) __________ attached to a carrier particle

ERYTHROCYTES

LATEX PARTICLES

GELATIN

SILICATES

BENTONITE

CHARCOAL

Carrier particles:

• ___________ - Always cross react with heterophile antibodies

• ______ __________ (common)

• __________

• _______

• ________

• _________

ANTISTREPTOLYSIN O (ASO) LATEX AGGLUTINATION

• The chemical particle that we have here is the latex particle, so the patient serum that contains antibodies will react with the latex particle then visible agglutination will occur.

• That means there is a particular antibody that could be present on the serum of the patient against Streptolysin O.

RHEUMATOID FACTOR (RF)

Seen in autoimmune disorder: Rheumatoid Arthritis

ANTINUCLEAR ANTIBODY (ANA) TEST

Screening test for Systemic Lupus Erythematosus

REVERSE PASSIVE AGGLUTINATION

ANTIGEN

ANTIBODY

(a)_________ __________ _____________

• Antibody is attached to a carrier particle

• Detects: (b)_________

• Reagent: (c)_________ attached to a carrier particle

REACTIVE

OUTWARD

The antibody must still be (a) _______ and is joined in such a manner that the active sites are facing (b)__________.

Fc

Fab

Carrier particles should have the ability to absorb the (a)___ region of the antibody so that the (b)____ region is exposed to the environment finding specific antigen.

C-REACTIVE PROTEIN (CRP) LATEX AGGLUTINATION TEST

Generarally a marker for acute inflammation.

AGGLUTINATION INHIBITION

PARTICULATE

SOLUBLE ANTIGENS

LIMITED ANTIBODY COMBINING SITES

AGGLUTINATION

POSITIVE

• (a)____________ _________ reactions are based on competition between (b)__________ (1st reagent) and (c)_______ ___________ (patient sample) for (d)________ __________ _________ ______ (2nd reagent), and a lack of (e)__________ is an indicator of a (f)__________ reaction.

ANTIGEN

ANTIBODY

Either (a) __________ or (b) ___________ can be attached to the particles

CLASSIC PREGNANCY TEST

POSITIVE RESULT

NEGATIVE RESULT

(a) __________ ____________ _____

• Antibody reagent and the patient antigen.

• (b) ___________ ________ in pt using the classic pt

- The reagent antibody will be bind to the patient antigen, they will be no attachment to the indicator cells therefore no agglutination reaction will be formed.

• (c) _________ _________

- The reagent antibody will not bind to the patient antigen, and the indicator cells will attach to the patient antibody which will form an agglutination reaction.

ABO TYPING USING SALIVA

____ __________ _______ ________

a. Specimen: Saliva

b. Principle: Hemagglutination Reaction

ANTI-SERA

PX SALIVA

KNOWN CELLS

(a)_____ ______ + (b)____ _____ (containing soluble antigens) + (c)_____ _______ (indicator cells)

POSITIVE

NEGATIVE

(a) ____ - No agglutination

(b) ____ - Agglutination

SECRETOR STATUS

Incorporated in some of the tests like the detector of (a) _________ __________.

(a)__________ _________ – how we able to identify the blood type of the specific individual using secretion using the saliva.

COAGGLUTINATION

__________________ detects antigen.

• Reverse passive agglutination but uses bacteria (Staphylococcus aureus) as the carrier particle.

• It is also the active site that faces outward and will have the capability to react with the specific antigen.

S. AUREUS

______________ is most frequently used, because it has a protein on its outer surface, Protein A.

PROTEIN A

________ - it naturally absorbs the Fragment crystallizable (FC portion) of the antibody molecule.

AHG-MEDIATED AGGLUTINATION

____-______________ ______________________

• Also known as Coomb’s Test

• Detects non agglutinating antibodies by means of coupling with a second antibody.

HYBRIDOMA TECHNIQUES

The key component of the test is antibody to human globulin that is made in animals (rabbit/mouse) or by means of __________ ______________.

RBC

Reason this process is performed: Is to detect if the RBC is sensitized with IgG or complement. We are dealing with sensitization of the _____ (in vivo/In vitro).

IgM ANTIBODIES

____ ____________ can bind to the corresponding antigen and it directly agglutinates.

DIRECT

INDIRECT

The Coombs’ test can be divided into two different types, _________ and ____________, each of which has a different purpose.

DIRECT

VIVO ATTACHMENT

(a)__________

• Used to demonstrarate direct in (b)______ ____________ of the antibody, usually IgG or complement to to an individual’s RBC.

• it is called “(a)________” because we are referring to the sensitization of the red cells that are tested directly as they come (directly) from the body.

INDIRECT

(a)____________

• In-Vitro

• Used to determine the presence of a particular antibody in a patient.

• Part of pre-transfusion protocols.