[M] Lec 02.3: Intro to Sero (Immunologic Reactions)

Question 1

Refers to

● Their basis is the primary Ag-Ab reaction.
● It measures the antigen binding capacity of antibodies.
● It measures low molecular weight antigen such as bovine serum albumin and hapten.

Answer 1

Primary Ag-Ab Reaction

Question 2

Classification

1. Farr Technique/Ammonium Sulfate Precipitation
2. Equilibrium Dialysis
3. Labeled Immunoassay: Radio Immunoassay (RIA), Enzyme Immunoassay (EIA), and Immunofluorescence
   Assay (A.K.A. Fluorescence Immunoassay).

Answer 2

Primary

Question 3

1. Specific
2. General

A. Enzyme Immunoassay (EIA)
B. Enzyme-Linked
Immunosorbent Assay (ELISA)

Answer 3

1. B
2. A

Question 4

T or F:

○ In the laboratory, we now use Labeled Immunoassays.
○ We rarely use Agglutination and Precipitation tests because Labeled Immunoassays are more sensitive.

Answer 4

True

Question 5

Arrange labeled immuno assays according to sensitivity (most to least)

Answer 5

1. RIA
2. EIA
3. IFA

Question 6

1. Use a precipitating reagent, particularly Ammonium Sulfate to visualize the reaction.
2. Use a dialysis device composed of two chambers—chamber A and chamber B— with a semipermeable membrane between them

A. Equilibrium dialysis
B. Farr Technique .

Answer 6

1. B
2. A

Question 7

1. Radio labels; beta or gamma counters
2. Enzyme labels; ELISA reader
3. Fluorescent labels; fluorescent microscopes

A. EIA
B. RIA
C. IFA

Answer 7

1. B
2. A
3. C

Question 8

Refers:

• Farr Technique and Equilibrium Dialysis are commonly
  performed to measure below molecular weight antigen— like HAPTEN AND BOVINE SERUM ALBUMIN.
• Bovine Serum Albumin + I131 → (I131BSA) + Anti-BSA → no visible reaction
  ○ Bovine Serum Albumin will be coupled to a Radioisotope (I131), forming a complex.
  ○ The complex, when added with the antibody will produce no visible reaction.

Answer 8

Farr Technique

Question 9

Important for precipation in Farr Technique

Answer 9

Ammonium sulfate

Question 10

Determines the antigen binding capacity of antibody in Farr technique

Answer 10

Amount of I131

Question 11

Refers:

• Reference method for FT4 (Free Thyroxine)
• A dialysis device is used, which is divided into two chambers:
  Chamber A and Chamber B.
  ○ Cellulose is often used as a semi-permeable membrane.
• During the initial state of testing, antigens are placed in Chamber B, while antibodies are placed in Chamber A

Answer 11

Equilibrium dialysis

Question 12

Refers to reactions that are visible

Answer 12

Secondary

Question 13

Classify the ff reactions:

1. Precipitation
2. Agglutination
3. Complement Fixation
4. Neutralization (In Vitro)

Answer 13

Secondary

Question 14

Arrane from sensitive to least sensitive serologic tests

Answer 14

1. Neutralization (In Vitro) - most sensitive
2. Complement Fixation
3. Agglutination
4. Precipitation - least sensitive

Question 15

Define the ff secondary serologic texts

1. The combination of SOLUBLE AG WITH AB to produce an INSOLUBLE COMPLEX.
2. The antigen involved is insoluble; has a stronger reaction compared to precipitation
3. Fixation of complement by antigen antibody (immune) complex.
4. When test serum is mixed with standard amount f toxin/ virus, there is decrease of toxicity of toxin or infectivity of virus.

Answer 15

1. Precipitation
2. Agglutination
3. Complement fixation
4. Neutralization

Question 16

Two types of precipitation medium

Answer 16

Liquid and semi-liquid gel

Question 17

1. Slide Precipitation, Tube Precipitation, Capillary Precipitation makes use of capillary tube, Interfacial ‘ring’ Test uses test tubes
2. Immunodiffusion, electroimmunodiffusion

Answer 17

1. Liquid
2. Semi-solid gel

Question 18

Refers to

• Non-treponemal/non-specific test for syphilis
  ○ It is non-specific because RPR only detects reagin.
• Principle: FLOCCULATION
• Detection of antibody called Reagin
• Uses Choline-chloride instead of heat inactivation. Choline-chloride is used to inactivate complement.

Answer 18

Slide precipitation (RPR)

Question 19

Study the procedures nalang

Answer 19

Pls

Question 20

Match (Slide precipitation)

1. Medium and large clumps
2. No clumping or very slight roughness
3. Small clumps

A. Weakly reactive
B. Non-reactive
C. Reactive

Answer 20

1. C
2. B
3. A

Question 21

Tests we can do in RPR

1. NOT exact amount of value since titer only relies on highest dilution with positive reaction
2. We only detect the presence or absence of the antigen or the antibody
3. Determines the exact amount by using machine/equipment

Answer 21

1. Semi-quantitative
2. Qualitative
3. Quantitative

Question 22

Qualitative RPR

1. SCREENING TEST and DETECTING ANTIBODY
2. CONFIRMATORY TEST and DETECTING ANTIGEN

Answer 22

1. Reactive/Non
2. Positive/Negative

Question 23

In semi-quantitative RPR, dilute specimens with what buffer?

Answer 23

Glycine NaCl

Question 24

Give me the CRP of the ff

1. 1+1 (1:2)
2. 1+3 (1:4)
3. 1+7 (1:8)
4. 1+15 (1:16)
5. 1+31 (1:32)

Answer 24

1. 12
2. 24
3. 48
4. 96
5. 192

Question 25

1. Best example is RPR 2. We can use centrifuge to speed up the reaction as compare to slide precipitation where we ise mechanical rotator 3. Can be performed using capillary tube or test tube A. Slide precipitation B. Ring precipitation C. Tube precipitation

Answer 25

1. A 2. C 3. B

Question 26

Define terms of precipitation 1. Refers to the reactant– the antigen or antibody 2. Refers to the direction of the migration 3. Direction of the migration is either upward or downward 4. Refers to radial movement (from center inner to outer) 5. Either the antigen is moving or the antibody is moving to meet to form precipitation 6. Both the antigen and antibody are moving to meet to form precipitation

Answer 26

1. Diffusion 2. Dimension 3. Single dimension 4. Double dimension 5. Single diffusion 6. Double diffusion

Question 27

Refers to: * example of precipitation reaction, therefore this is classified under secondary Ag-Ab reaction * Precipitation reactions that is why these tests are not commonly performed in laboratory and are obsolete and usually needed for board exam purposes only

Answer 27

Immunodiffusion tests

Question 28

Factors that can affect immunodiffusion tests

Answer 28

○ Size of particles ○ Size of antigen ○ Temperature of the reaction ○ Viscosity of the gel ○ Amount of hydration

Question 29

Match 1. Only targeted antigen is moving, direction of movement is radially 2. Only antigen is moving 3. Both the antigen and antibody is moving 4. Antigens B and C are reactive with the patient's serum in the center well A. Single Diffusion, Single Dimension (Oudin test) B. Single Diffusion, Double Dimension (Radial Immunodiffusion) C. Double Diffusion, Single Dimension (Modified Oudin with Oakley & Fulthorpe) D. Double Diffusion, Double Dimension (Ouchterlony Technique)

Answer 29

1. B 2. A 3. C 4. D

Question 30

Two methods in single diffusion, double dimension

Answer 30

1. Fahray 2. Mancini

Question 31

Refers to * Precipitation reaction * Classified under secondary antigen antibody reaction * Also known as Immunoelectrophoresis * Electrophoresis’ purpose is to enhance the mobility of the reactants. To increase the movement and mobility of the antigen and antibody towards one another

Answer 31

Electroimmunodiffusion Tests

Question 32

Electroimmunodiffusion Tests 1. Also known as Laurel Immunoelectrophoresis or Rocket electroimmunodiffusion; Based on single electroimmunodiffusion so there is only one reactant moving 2. Also known as countercurrent immunoelectrophoresis or cross immunoelectrophoresis; Both antigen and antibodies are moving. 3. Counterpart of the radial immunodiffusion; Only one reactant is moving and is double dimension 4. Developed by Grabar and Williams; Both antigen and antibodies are moving; Double dimension means the direction or movement is radially.

Answer 32

1. Single Electroimmunodiffusion, Single dimension 2. Double Electroimmunodiffusion, Single Dimension 3. Single Electroimmunodiffusion, Double dimension 4. Double Electroimmunodiffusion, Double dimension

Question 33

1. The antibody/ anti-serum is placed in the trough 2. The antibody/ anti-serum is placed directly on thegel’s surface (agar) A. Immunofixation B. Immunoelectrophoresis

Answer 33

1. B 2. A

Question 34

Types of agglutination

Answer 34

1. Direct Agglutination 2. Indirect/Passive Agglutination 3. Reverse Passive Agglutination 4. Agglutination Inhibition 5. Hemagglutination

Question 35

Clinical application of agglutination

Answer 35

* Coombs’ Test * Bentonite Test * Latex Fixation Test * Rose Waaler Test

Question 36

Types of agglutination 1. Agglutination of natural antigen 2. Antibody is artificially attached to a particulate carrier 3. Uses antigen that is artificially attached to a particulate carrier 4. Agglutination of RBC due to antibody (e.g. in vivo sensitization), viruses, bacteria, or other biologic substance. 5. Based on Blocking Principle; The homologous antigen inhibits the agglutination of antigen coated particle

Answer 36

1. Direct 2. Reverse 3. Indirect 4. Hemagglutination 5. Agglutination Inhibition

Question 37

Uses of direct agglutination

Answer 37

1. Detection of antigen 2. Detection of antibody

Question 38

HCG is an example of which type of agglutination

Answer 38

Agglutination inhibition

Question 39

Types of hemagglutination 1. Positive result (+): Carpet/Mat cells or Hemagglutination 2. Pased on blocking principle 3. Useful in the diagnosis of Influenza and mumps 4. Useful in the diagnosis of influenza, mumps, and German measles 5. Involved are natural antigens

Answer 39

1. Indirect 2. Hemagglutination inhibition 3. Virus inhibition 4. Virus hemagglutination inibition 5. Direct hemagglutination

Question 40

Two systems involved in complement fixation

Answer 40

○ Test system/Bacteriolytic system ○ Indicator system/Hemolytic system

Question 41

Main components of complement fixation

Answer 41

1. Known Ag reagent (ex. beef heart extract, bacterial Ag) 2. Complement from Guinea pig serum (Best Source) 3. Hemolysin or Amboreceptor 4. Indicator cells (Sensitized Sheep RBCs)

Question 42

Best source of complenent

Answer 42

Guinea Pig Serum

Question 43

Complement fixation 1. Positive result 2. Negative result A. No hemolysis B. Hemolysis

Answer 43

1, A 2. B

Question 44

Types of toxin neutralization 1. Tertiary antigen-antibody reaction 2. Secondary antigen-antibody reaction

Answer 44

1. In vivo 2. In vitro

Question 45

Types of virus neutralization

Answer 45

1. Pock Reduction Test (In Ovo) 2. Plaque Reduction Test 3. Metabolic Inhibition Test 4. Tissue Culture Technique