immunology lab 3 quiz

test method hemagglutination description

aggregation of red cells due to a specific antibody cross-linking the antigens

hemagglutination example

blood typing

test method precipitation description

antigen-antibody reaction where both are soluble, forms a complex that precipitates

precipitation example

immunodiffusion

test method flocculation description

a precipitation test that uses charcoal to visualize the antibody-antigen reactionfl

flocculation example

rapid plasma reagin

test method latex agglutination description

aggregation of latex particles due to a specific antigen or antibody cross-linking the latex

latex agglutination examples

rheumatoid factor & CRP

test method capture assay description

creates an antibody-antigen complex, with one of them bound to a color conjugate

capture assay examples

antigen capture: SureVue HCG, Immunosorbent: Mono

test method indirect immunofluorescence description

detect antinuclear antibodies using Hep-2 cells as a substrate and a secondary antibody with fluorescent conjugate

indirect immunofluorescence example

antinuclear antibody testing

soluble

secreted and free floating in solutioni

insoluble

bound and cannot be free floating in solution

for the hemagglutination test method, which is soluble and insoluble

antigen is insoluble and antibody is soluble

for precipitation test method, which is soluble and insoluble

both the antigen and antibody are soluble

for latex agglutination test method, which is soluble and insoluble

we choose, whatever is bound to latex is insoluble

agglutination is characterized by

macroscopic clumping due to the repeated bonds formed by antibodies and twheir corresponding insoluble antigen

what do antigen-antibody molecules form in agglutination

lattice

what happens to the IgM antibodies in immunoglobulin agglutination

the antibodies will agglutinate immediately. multivalent (pentamer), cold agglutinin

what happens to IgG antibodies in immunoglobulin agglutination

the antibodies require an additional test to detect. bivalent. agglutinates better in body temp

variables in antigen-antibody reactions

ionic strength, temperature, incubation time, centrifugation, antigen-antibody concentration, affinity, avidity

affinity

strength of one antibody-antigen

does IgG have high affinity or avidity

affinity

aVidity

strength of Various antigen-antibodies

does IgM have high affinity or avidity

avidity

3 zones for antigen-antibody concentration in order of left to right in a graph

antibody excess zone, equivalence zone, antigen zone (negative parabola graph)

free antibody concentration left to right in graph across the 3 stages

+--

free antigen concentration left to right in graph across the 3 stages

- - +

x variable in antigen-antibody concentraiton graph

antigen added

y variable in antigen-antibody concentration graph

immune complex precipitated

prozone/postzone problems cause what issue?

false negatives

procedure part a

label 12 tubes numerically

transfer 2 drops of sample serum to the tubes

add 1 drop of red cells

mix and centrifuge for 20 seconds

gently shake and interpret agglutination strength, record results

lowest agglutination grade

+1

4+ agglutination grade means

one solid clump

3+ grade means

several LARGE clumps

2+ grade means

numerous SMALLER clumps

1+ grade

barely discernable clumps

goal of serial dilutions

obtain a titer as an indication of the concentraiton of the antibody

titer

concentration of a solution as determined by titration

what 3 things are serial dilutions used for

determine immunity

look at rise and fall of antibody level to either evaluate prognosis or determine stage of disease

solve a prozone problem. dilution of antibody prevents false negatives

diluent

material used to prepare the dilutionw

what is the diluent in our lab?

normal saline/physiological/0.9%

endpoint or titer

reciprocal of last positive dilution

in which tubes does the saline go?

tubes 2-10

in which tubes does the sample serum go

tubes 1-2

transfer 100 nanoliters from tube 2 until when

until tube 10. discard last 100 nanoltiers

add 100 nanoliters of red cells to which tubes and when

tubes 1-10. after the dilution has been prepared

what is in the sharps for disposal?

glass test tubes generated AND plastic pipette tips

what goes in biohazard for disposal

soft plastic transfer pipettes AND paper towels that touched serum or blood