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Immunology and Serology (Quiz 2) > Precipitation Methods > Flashcards

Precipitation Methods Flashcards

1
Q

Principle: Soluble antigen combines with soluble antibody to produce visible insoluble complexes

A

Precipitation

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2
Q

Clumping together of particles to form visible masses over a narrow range of antigen concentration

A

Flocculation

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3
Q

Principle: Soluble antigens react with specific antibodies to form a precipitate of fine particles

A

Flocculation

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4
Q

Applications of Flocculation

A

Venereal Disease Research Laboratory (VDRL) tests

Rapid Plasma Reagin (RPR)

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5
Q

Principle: Light scattering by immune complexes is measured
● scattering of light is proportional to the size and amount of immune complexes formed

A

Nephelometry

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6
Q

Applications of Nephelometry

A

Immunoglobulins

Complement

C-reactive protein

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7
Q

Measures the decrease in light intensity in a solution containing immune complexes

A

Turbidimetry

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8
Q

Measures the reduction of light transmitted at 180 ° angle.

A

Turbidimetry

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9
Q

Measures transmitted light at 90 ° angle

A

Nephelometry

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10
Q

if only one reactant (usually antigen) is moving

A

Single Diffusion

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11
Q

if both antigen and antibody are moving through the medium

A

Double diffusion

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12
Q

if the reaction in a medium have only one effective dimension for antigen and antibody migration (i.e., up and down)

A

Single dimension

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13
Q

if the reaction is in circular holes (i.e., wells) cut in a gel on a flat surface, diffuses from the wells radially

A

Double dimension

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14
Q

Principle: Known Antibody fixed in agar + Unknown Antigen (overlaid) → Precipitin lines

A

Single Linear Diffusion (SLD) or Oudin Technique

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15
Q

Applications of Single Linear Diffusion (SLD) or Oudin Technique

A

Detects multiple antigen- antibody reactions

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16
Q

Principle: Known Antibody fixed in agar + Unknown Antigen (well cut in agar plate) → Precipitin ring

A

Single Radial diffusion

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17
Q

2 types of Single Radial diffusion

A

Fahey Method & Mancini Method

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18
Q

diameter of precipitin ring at 24 hours (Read before it reaches the maximum at 6-12 hours)

A

Fahey Method

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19
Q

area of precipitin ring formed at 48 hours.

A

Mancini Method

20
Q

Uses antibody not in excess.

A

Fahey method

21
Q

● Diffusion time for result reading is critical.
● Diffusion endpoints may not be reached,
affecting accuracy.

A

Fahey method

22
Q

Uses excess antibody in the gel for higher
sensitivity and accuracy.

A

Mancini method

23
Q

More commonly used in commercial
immunodiffusion plates.

A

Mancini method

24
Q

Principle: Antigen diffuses out of well in a gel containing antibody → Precipitin ring forms → Diameter proportional to the concentration of antigen

A

Radial Immunodiffusion (RID)

25
Q

technique used to measure antigen concentration based on the formation of a lattice network between ag and ab in a gel.

A

Radial Immunodiffusion (RID)

26
Q

2 types of Radial Immunodiffusion (RID)

A

Endpoint Method (Mancini Method)

Kinetic Method (Fahey Method)

27
Q

● Ring diameter readings are taken at about 19 hours before equivalence is reached.

● The diameter is proportional to the log of the concentration.

A

Kinetic Method (Fahey Method)

28
Q

● Antigen is allowed to diffuse to completion until equivalence is reached, indicated by no further change in ring diameter.

● The square of the diameter is directly proportional to the antigen concentration.

A

Endpoint Method (Mancini Method)

29
Q

Applications of Radial Immunodiffusion (RID)

A

Immunoglobulins, complement

30
Q

No longer commonly performed except for low-volume testing of IgD and IgG

A

Radial Immunodiffusion (RID)

31
Q

Principle: Antigens and antibodies diffuse out from wells cut in gel and precipitin lines where they meet

A

Ouchterlony technique / double immunodiffusion

32
Q

Ouchterlony technique / double immunodiffusion

Three basic reaction patterns:

A

Identity
Non-identity
Partial identity

33
Q

a single smooth arc of precipitation forms between the antigens and antibodies

A

Identity

34
Q

two separate lines of precipitation cross each other

A

Non-identity

35
Q

two precipitating lines meet, forming a
spur

A

Partial identity

36
Q

Applications of Ouchterlony technique / double immunodiffusion

A

Fungal antigens
Extractable nuclear antigens

37
Q

Principle: Antigens and antibodies are placed in wells that are directly opposite one another in a gel → an electrophoretic charge is applied to drive the reactants toward each other → precipitin band forms where they meet

A

Countercurrent immunoelectroph oresis (CIE)

38
Q

● To rapidly check any antisera for the presence and specificity of antibodies for a particular antigen.

● To detect antigens and/or antibodies in serum for diagnosis of a particular disease

A

Countercurrent immunoelectroph oresis (CIE)

39
Q

Applications of Countercurrent immunoelectroph oresis (CIE)

A

Bacterial antigens

40
Q

Principle: Proteins are separated by electrophoresis, then subjected to double diffusion with reagent antibodies placed in a trough cut in the agar → shape, intensity, and location of the precipitin arcs developed are compared with those of a normal control

A

Immunoelectrophoresis (IEP)

41
Q

Applications of Immunoelectrop horesis (IEP)

A

Serum proteins, including immunoglobulins

42
Q

Principle: Proteins are separated by electrophoresis → cellulose acetate strip impregnated with antiserum is placed on the separated proteins → the antiserum diffuses into the gel, and antigen-antibody complexes precipitate

A

Immunofixation electrophoresis (IFE)

43
Q

Gel diffusion + Electrophoresis

A

Immunoelectrop horesis (IEP)

44
Q

Protein electrophoresis + immunoprecipitation

A

Immunofixation electrophoresis (IFE)

45
Q

Applications of Immunofixation electrophoresis (IFE)

A

Identification of immunoglobulin s in monoclonal gammopathies, Bence-Jones proteins

46
Q

Principle: An electrical charge is applied to a RID assay → the height of the rocket-shaped precipitin band is proportional to the concentration of antigen

A

Rocket Electrophoresis

47
Q

Applications of Rocket Electrophoresis

A

Immunoglobulins, complement, alpha-fetoprotein

Immunology and Serology (Quiz 2) (2 decks)

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