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FA1 Immunology > Lecture 22 11/9/23 > Flashcards

Lecture 22 11/9/23 Flashcards

1
Q

What is an immunological assay?

A

test that makes use of antigen-antibody interactions

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2
Q

What are the clinical applications of immunological tests?

A

-confirming diagnosis of infectious disease
-assess patient’s immune status
-determine exposure to a pathogen
-assess immunity in response to a vaccine

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3
Q

What is the role of a qualitative test?

A

determine if a particular substance is present in the clinical sample

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4
Q

What is the role of a quantitative test?

A

assess how much of the tested antigen/antibody is present

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5
Q

What is a “stat” test?

A

-test with a quick turnaround time
-ordered when result is needed quickly

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6
Q

What is a “routine” test?

A

-test with a less immediate turnaround for patients with less of a need for immediate results
-results available within hours to days

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7
Q

Which section of the antibody is labelled/tagged in immunological assays?

A

the Fc segment

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8
Q

What is an enzyme label?

A

-enzyme will cleave a substrate
-cleavage produces a detectable product (light, color, etc)

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9
Q

What is a fluorescent label?

A

-fluorophore that absorbs light at a specific wavelength
-emits light at a different wavelength

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10
Q

What is a colored bead label?

A

-large synthetic particle
-color can be read visually when enough aggregate together

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11
Q

What are the characteristics of ELISA assays?

A

-performed in 96-well plate
-use enzyme labels (horseradish peroxidase)
-read using spectrophotometer
-color change is positive
-can be qualitative or quantitative

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12
Q

What are the steps of a direct ELISA?

A

-patient specimen fixed
-antigen-specific antibody with enzyme label applied
-unbound antibodies rinsed off after incubation
-substrate added; if cleaved by enzyme label, color change occurs

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13
Q

What is measured with a direct ELISA?

A

antigen within a patient specimen

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14
Q

What is measured with an indirect ELISA?

A

antibody within a patient specimen

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15
Q

What are the steps of an indirect ELISA?

A

-purified known antigen attached to solid phase
-patient serum added
-unbound antibodies washed off after incubation
-enzyme-labeled secondary antibody added
-unbound antibodies washed off after incubation
-substrate added; color change indicates cleavage by enzyme/positive

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16
Q

What are the characteristics of immunofluorescence assays?

A

-use fluorescent label
-read with fluorescence microscope
-performed on glass slides with 8-12 samples
-fluorescence is positive indicator
-can be qualitative or quantitative

17
Q

What are the characteristics of a direct fluorescence antibody assay?

A

-detects viral antigens in a specimen
-patient specimen fixed to slide
-fluorescent labelled antibodies added to specimen

18
Q

What are the characteristics of an indirect fluorescence antibody assay?

A

-detects antibodies that a patient has formed against a virus
-known viral antigens fixed to slide
-patient serum added
-fluorescently labeled antibodies added that attach to patient antibodies

19
Q

What is the role of serum protein electrophoresis?

A

separate and quantify different protein fractions in blood serum

20
Q

What is the gamma globulin fraction mostly made up of?

A

immunoglobulins (antibodies)

21
Q

What is hypergammaglobulinemia?

A

abnormal increase in antibody synthesis

22
Q

What are the two types of hypergammaglobulinemia?

A

-monoclonal
-polyclonal

23
Q

What are the characteristics of monoclonal gammopathies?

A

-increase in a single type of antibody
-associated with immunoglobulin-secreting cancers/myelomas
-monoclonal peak has narrow base with peak that is as sharp/sharper than albumin peak

24
Q

What are the characteristics of polyclonal gammopathies?

A

-increase in all major antibody types
-result from long term immune stimulation
-globulin peak has a broad and diffuse base

25
Q

What is flow cytometry?

A

technique used to detect and measure physical and chemical characteristics of a population of cells

26
Q

What is forward scattered light (FSC) proportional to?

A

cell-surface area or size

27
Q

What is side-scattered light (SSC) proportional to?

A

cell granularity/internal complexity

28
Q

Which conditions warrant the use of a flowcytometer?

A

-differentiation between B and T cell lymphoid neoplasms
-differentiation between reactive and neoplastic expansions of lymphocytes
-identification of specific subsets of lymphocytes
-prognosis in lymphoma

FA1 Immunology (24 decks)

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