Immunology- Labeled Assays
An additional advantage of an indirect assay is that
- it amplifies the reaction and makes the assay more sensitive -it allows for the determination of the class of the antibody that is bound
An indirect radioimmunoassay uses a different second antibody-binding step because of the radiolabel, but the same antibody binding step as
-an indirect enzyme immunoassay -an indirect fluorescent immunoassay -a capture assay
The presence of human heterophilic antibody is
-difficult to predict -the result of the patients interactions with animals of the species used to make the antibody in the assay -can cause false positive reactions in capture assays
HAMA
-is a human anti-mouse antibody -can cause false positives in capture immunoassays
In a sandwhich assay
-the antibody is sandwiched between a capture antigen and a labeled antigen -the antigen is sandwiched between a capture antibody and a labeled antibody
In a capture assay for IgM that reacts with rubella, an antibody human IgM binds the patient IgM, rubella antigen from the kit is added, and then an enzyme labeled antibody to rubella is added. What would happen to this assay if the patient had rheumatoid factor but no IgM antibodies to rubella?
The patient would appear positive for IgM rubella antibodies unless human IgG -that did not react with rubella was added as a specific blocker
Optical immunoassays are based on
a change in the color of light reflected from the surface when ag and ab are bound
For a small antigen with one epitope that is in the fluid phase, the best assay would be
a competitive assay
Which is a homogeneous assay?
a fluorescence polarization immunoassay
Which assay is the most sensitive?
a heterogenous labeled immunoassay
A fluorescent polarization assay is
a homogeneous labeled immunoassay
To measure the amount of a 200,000 MW tumor antigen in a patient's blood, the best assay is
a sandwich immunoassay
In a competitive immunoassay, the
amount of signal is inversely proportional to the amount of patient's antibody and can be determined from a standard curve
An indirect radioimmunoassay uses a different second antibody binding step because of the radiolabel but the same first antibody binding step as
an indirect enzyme immunoassay and an indirect fluorescent immunoassay
What two labeled immunoassays do not require instrumentation?
colloid immunoassay and optical immunoassay
In analysis of multiplex bead
one-color fluorescence is measured from the indirect immunoassay signal and one blended color is measured from the bead
Two assays used for point of care tests are
optical and colloid immunoassays
The first labeled assays developed were
radioimmunoassays
A western blot differs from a recombinant immunoblot because
the antigens on the nitrocellulose strip in the western blot have been electrophoresed and then blotted on to the blot and the recombinant immunoblot the antigens are just pipetted onto the blot
The assay that requires a trigger to measure the signal
the chemiluminescent assay
The kit for homogenous assay for meth comes with meth linked to G6P dehydrogenase, and antibody to the drug blocks the action of the enzyme when the patient's serum contains
the drug and there is more enzyme reactivity
To remove the effect of a patient's rheumatoid factor
the patients sample could be diluted in a buffer containing IgG that does not react with the test antigen
You have a patient who has rheumatoid factor. If you are performing an assay for various infectious diseases in which you capture the patients IgM, add the antigen, and then add a labeled antibody to the antigen....
the reactions with all antigens tested would be positive