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1. Imse > Immuno Part 2 Lec > Flashcards

Immuno Part 2 Lec Flashcards

1
Q

Immunoassays have been developed to detect either antigen or
antibody, and they vary from easily performed manual tests to
highly complex automated assays.

A

Precipitation Reaction

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2
Q

involves combining soluble antigen with soluble antibody to
produce insoluble complexes that are visible.

A

Precipitation

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3
Q

is the process by which particulate antigens aggregate to form
larger complexes when a specific antibody is present.

A

Agglutination

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4
Q

is the initial force of attraction that exists between a
single Fab site on an antibody molecule and a single epitope or
determinant site on the corresponding antigen.

A

Affinity

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5
Q

School-based laboratory manual test

A

Precipitation reaction

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6
Q

Most efficent immmunoglobulin in this type of reaction because of its small molecules which is much efficient in Precipitation reaction

A

IgG

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7
Q

indicator for agglutination reaction

A

Rbc

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8
Q

most efficient; antibody involved in large agglutination reaction. Has 10 valence

A

IgM

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9
Q

Large complex of agglutination

A

Clumping

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10
Q

Antibodies are capable of reacting with antigens that are structurally similar to the original antigen that induced antibody production.

A

Cross-reactivity

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11
Q

It is inline to affinity

A

Cross-reactivity

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12
Q

initial force of attraction of antigen and antibody is called

A

Cross-reactivity

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13
Q

represents the sum of all the attractive forces between an antigen and an antibody.

A

Avidity

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14
Q

involves the strength with which a multivalent antibody binds a multivalent antigen, and it is a measure of the overall stability of an antigen–antibody complex

A

Avidity

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15
Q

is essential to detecting the presence of an unknown, whether it is antigen or antibody

A

Avidity

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16
Q

A high avidity can actually compensate for a ______ (low or high) affinity.

A

Low

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17
Q

stabilization ng antigen-antibody complex binding

A

Avidity

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18
Q

antigen-antibody binding

A

Affinity

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19
Q

Precipitin curve, we are dealing with the

A

Zone of equivalence

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20
Q

Zone of equivalence where it is inlined with the

A

Prozone and postzone

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21
Q

the optimum precipitant curve in which the number of the multivalent sites of the antigen and antibody are approximately equal

A

Zone of equivalence

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22
Q

phenomenon wherein we are dealing with antibody excess.

A

Prozone

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23
Q

The antigen combines with only 1 or 2 antibody molecules and no cross linkages are formed

A

Prozone

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24
Q

antigen excess. No lattice network formed.

A

Postzone

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25
Q

Every available antibody site is bound with single antigen and no crosslinks are formed

A

Postzon

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26
Q

Measurement of precipitation by

A

light scattering

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27
Q

Determination of Precipitation via

A

Passive immunodiffusion techniques
Immuno electrophoretic techniques

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28
Q

Light scattering test involves

A

Turbidimetry
Nephelometry

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29
Q

Passive immunodiffusion techniques test involve

A

Radial immunodiffusion
Ouchterlony double diffusion/immunodiffusion

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30
Q

Immunoelectrophoretic technique test involved

A

Rocket immunoelectrophoresis
Immunoelectrophoresis
Immunofixation electrophoresis
Countercurrent immunoelectrophoresis

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31
Q

is a measure of the turbidity or cloudiness of a solution.

A

Turbidimetry

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32
Q

A detection device is placed in direct line with the incident light.

Collecting light after it has passed through the solution.

A

Turbidimetry

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33
Q

turbidimetry has the intensity of light transmitted thru the medium, so that the unscattered light is measured at _______ degrees angle from the incident light beam

A

180 degrees

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34
Q

the intensity of scattered light is measured. It is usually not necessarily at right angles to the incident light beam

A

Nephelometry

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35
Q

It can detect antigen or antibody but we usually use antibody as reagent and the patient sample to detect the unknown antigen

A

Nephelometry

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36
Q

Nephelometers measure light scatter at angles ranging from ______degrees to about ______degrees.

A

10 degrees to 90 degrees

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37
Q

This would be the reaction allowed to run essentially to completion
which means that large particles tend to fall out of solution and decreases the amount of the scatter

A

End point nephelometry

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38
Q

provides accurate and precise quantitation of serum proteins, and due to automation, the cost per test is typically lower than other methods.

A

nephelometry

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39
Q

Nephelometry Can also be used as a quantitation of immunoglobulins such as _____, ___, and _____, ______ even the Kappa and Lambda light chain and it is exclusively done by nephelometry.

A

IgG, IgA, and IgM, IgE

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40
Q

Order of nephelometry device

A

light source→ monochromator → cuvette → lens → detector

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41
Q

The precipitation of antigen–antibody complexes can also be determined in a support medium such as a ______

A

Gel

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42
Q

Passive immunodiffusion techniques A high-molecular-weight complex polysaccharide derived from __________, and ___________, a___________, are used for this purpose.

A

seaweed, agarose and purified agar

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43
Q

The precipitation of antigen–antibody complexes can also be determined in a support medium such as a gel.

A

Passive immunodiffusion technique

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44
Q

The rate of diffusion is affected by the size of the particles, the temperature, the gel viscosity, and the amount of hydration.

A

Passive immunodiffusion techniques

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45
Q

involves the stabilization and visualization

A

Agar or agarose gel

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46
Q

A modification of the single-diffusion technique was the

A

Radial immunodiffusion

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47
Q

who was the first to use gels for precipitation reactions, and he pioneered the technique known as single diffusion.

A

James Oudin

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48
Q

Radial immunodiffusion has
positive result would be the formation of the

A

precipative bonds

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49
Q

Radial Immunodiffusion is also called as

A

RID

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50
Q

Antibody is uniformly distributed in the support gel, and antigen is applied to a well cut into the gel.

A

RID

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51
Q

The area of the ring obtained is a measure of antigen concentration, and this can be compared to a standard curve obtained by using antigens of known concentration.

A

RID

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52
Q

Two techniques for measurement of RID

A

Mancini method

Fahey and Mckelvey method

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53
Q

DIAMETER OF THE RING = antigen concentration

A

Mancini method

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54
Q

Mancini method occurs between _____ and ______ hours

A

24 and 72 hours

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55
Q

Mancini method determines

A

IgM and IgG

56
Q

Determination of IgM:

A

completed at 50-72hrs

57
Q

Mancini method forms:

A

Ring formation

58
Q

Determination of IgG:

A

Complete at 24 hours

59
Q

Mancini method involve in method of

A

End point method

60
Q

the diameter is proportional to the log of the concentration

A

Fahey and mckelvey method

61
Q

Fahey and mckelvey method involves method We are using measurement taken before the point of equivalence is reached

A

Kinetic method

62
Q

Fahey and mckelvey methods Readings is taken for about

A

18 hours

63
Q

Sources of error include in radial immunodiffusion

A

● overfilling or under filling the wells,
● nicking the side
● spilling sample
● improper incubation time and temperature,
● incorrect measurement.

64
Q

Radial immunodiffusion has been used to measure

A

IgG, IgM, IgA, and complement components.

65
Q

In this technique, both antigen and antibody diffuse independently through a semisolid medium in two dimensions, horizontally and vertically.

A

Ouchterlony double diffusion

66
Q

has a smooth curve which means it have the same antigen that is present

A

Category A

67
Q

demonstrates a cross line pattern which shows two separate reactions and indicates that the compared antigens share no common epitotes

A

Category B

68
Q

there’s a fusion of 2 lines with a spur indicating partial identity.

A

Category C

69
Q

Also called as ‘Laurel Technique” because it was developed by Laurel in early 1960s

A

Rocket immunoelectrophoresis

70
Q

Rocket immunoelectrophoresis Also called as ‘_____________” because it was developed by Laurel in early 1960s

A

Laurel Technique

71
Q

The end result of rocket immunoelectrophoresis

A

Conical line

72
Q

Double-diffusion technique that incorporates electrophoresis current to enhance results.

A

Immunoelectrophoresis

73
Q

Immunoelectrophoresis Introduced by __________ and __________ in 1953, this is performed as a two-major step process and can be used for semi quantitation of a wide range of antigens.

A

Grabar and Williams

74
Q

this is performed as a two-major step process and can be used for semi quantitation of a wide range of antigens.

A

Immunoelectrophoresis

75
Q

2 Step process of immunoelectrophoresis

A

Electrophoresis

Diffusion of antigen and antibodies

76
Q

Immunoelectrophoresis Reaction takes place within_________

A

18-24hrs.

77
Q

procedure has been used as screening tool for the differentiation of many serum proteins including the major classes of immunoglobulins.

A

Immunoelectrophoresis

78
Q

Immunoelectrophoresis used in the laboraratory for the determination of

A

Myelomas

79
Q

Immunoelectrophoresis can detect

A

waldenstroms
macroglobulinemia
malignant lymphomas
other lymphoproliferative disorders

80
Q

causes increased bacterial infection

A

IgG deficiency

81
Q

Either increase value of antigen or deficient

A

B lymphocyte deficiency

82
Q

as first described by Alper and Johnson.

A

Immunofixation electrophoresis

83
Q

Similar to immunoelectrophoresis

A

Immunofixation electrophoresis

84
Q

Immunofixation electrophoresis meduim can be use

A

Agarose
Cellulose acetate

85
Q

Immunofixation electrophoresis Reaction Time:

A

Less than 1 hour

86
Q

Immunodiffusion takes place in a shorter time and results in a higher resolution than when antibody diffuses from a trough

A

Immunofixation electrophoresis

87
Q

Modification of immunoelectrophoresis.

A

Countercurrent immunoelectrophoresis

88
Q

Based on the movement of antigen towards the
anode and of the antibody towards the cathode during the passage of the electric current through agar.

A

Countercurrent immunoelectrophoresis

89
Q

Countercurrent immunoelectrophoresis reaction to take place

A

30-60 mins

90
Q

are designed for antigens and antibodies that may be small in size or present in very low concentrations.

A

Label immunoassay

91
Q

The presence of such antigens or antibodies is determined indirectly by using a

A

Labeled reactant

92
Q

The substance to be measured is known as the

A

Analyte

93
Q

On this Labelled immunoassays here are some examples of analytes:

A

Bacterial antigen hormone
Drugs ( test kits)
Tumor markers
Specific immunoglobulins

94
Q

is an antibody bonded to an analyte.

A

Labelled immunoassay

95
Q

Labeled immunoassays have made possible rapid quantitative measurement of many important entities such as

A

Viral antigen

96
Q

All the test kits that we encounter in immunology and serology are examples of Labelled immunoassays:

A

Mallaria test kits
Dengue NS1 for detecting recent dengue
infections
Dengue Duo for detecting IgM or IgG
Pregnancy test kits

97
Q

Characteristic of labelled immunoassay

A

o Competitive Assay
o Non-Competitive Assay

98
Q

Classification of labelled immunoassays

A

o Radioimmunoassay (RIA)
o Enzyme Immunoassay (EIA) o Fluorescent Immunoassay

99
Q

All the reactants are mixed together simultaneously , and a labelled antigen competes with unlabeled patient antigen for a limited number of antibody-binding sites

A

Competitive immunoassay

100
Q

Competitive immunoassay The amount of bound label is ______________ to the concentration of the patient antigen.

A

Inversely Proportional

101
Q

Antibody, is first passively absorbed to a solid phase. (in solid phase we use wells)

A

Non-competitive immunoassay

102
Q

Non-competitive immunoassay The amount of label measured is _________ to the amount of patient antigen

A

Directly proportional

103
Q

Non-competitive immunoassay is often called as

A

Captured antibody

104
Q

discovered by Georges Kohler and Cesar Milsten

A

Monoclonal antibodies

105
Q

remains a constant source of highly specific antibody

A

Monoclonal antibodies

106
Q

Antibodies in the Labelled immunoassays are characterized as

A

Monoclonal antibodies

107
Q

Monoclonal antibodies aka

A

Calibrators

108
Q

Are unlabeled analytes that are made up in known
concentrations of the substance to be measured

A

Calibrators

109
Q

Most immunoassays used the change in absorbance measured by

A

Spectrophotometry

110
Q

Quality Controls: should be run.

A

blank, negative control, and high and low positive control

111
Q

Pioneered by Rosalyn Yalow and Solomon Berson.

A

Radioimmunoassay

112
Q

Used radioactive substances as a label

A

Radio immunoassay

113
Q

Most common Label for RIA is Iodine

A

125I (Iodine 125), 131I (Iodine 131), 3H (tritiated Hydrogen)

114
Q

was originally based on the principle of competitive binding labelled antigen competes with unlabelled patient antigen for a limited number of antibody-binding sites.

A

Radioimmunoassay

115
Q

Ria The amount of bound label is ______________ to the concentration of the labelled antigen.

A

Inversely Proportional

116
Q

Advantages in using enzymes as label

A

cheap and readily available
Long shelf-life
easily adapted
Easy to dispose
little reagent is necessary

117
Q

Common enzymes used in EIA:

A

Horseradish peroxidase (MOST COMMON)
ALP (MOST COMMON)
G-6-PD
Beta-D-galactosidase

118
Q

Enzyme-labeled antigen competes with unlabeled patient antigen for a limited number of binding sites on antibody molecules
that are attached to a solid phase

A

Competitive Enzyme immuno assay

119
Q

Similar to RIA

A

Enzyme immunoassay

120
Q

Ezyme immunoassay aka

A

Direct elisa

121
Q

referred to as indirect enzyme-linked immunosorbent assays (ELISA), because the enzyme-labeled reagent does not participate in
the initial antigen–antibody binding reaction.

A

Non-competitive enzyme immunoassay

122
Q

Non-competitive EIA Enzymatic activity

A

Direct proportional

123
Q

Membrane-based cassette assays are a relatively new type of enzyme immunoassay

A

Rapid immuno assay

124
Q

The analyte is applied at one end of the strip and migrates toward the distal end, where there is an absorbent pad to maintain a constant capillary flow rate.

A

Immunochromatography

125
Q

Cassette test 2 lines indicates

A

Positive results

126
Q

1 line on control (C) means

A

Negative results

127
Q

1 line on test (T) or no lines at all indicates

A

Invalid results

128
Q

A form of immunoassay that uses label that fluoresce upon interaction with the desired analytes.

A

Fluorescent immunoassay

129
Q

these fluorescent compounds are called

A

fluorophores or fluorochromes

130
Q

fluorophores or fluorochromes compound

A

rhodamine, fluorescein, phycoerythrin

131
Q

have a red-orange fluorescence

A

Phycoerythrin

132
Q

Most commonly used fluorophores/fluorochromes:

A

Fluorescein

133
Q

→ absorbs maximally at 490 to 495 nm and emits a “green color” at 517-520 nm

A

Fluorescein

134
Q

absorbs at 550 nm and emits “ red colors” at 580 to 585 nm

A

Tetramethylrhodamine

135
Q

there is a known antigen is fixed on the slide. This is for the antigen detection only.

A

Direct IFA

136
Q

there is a known antigen fixed on the slide and it is used both for antigen and antibody detection. It involves a two step process.

A

Indirect IFA

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