working conference 1

presence of Ab in pt's sera

• 1. Ag coated onto surface of plastic well
• 2. pt's serum (w/ Ab) added to well; binding of Ab/Ag and excess washed away
• 3. enz-labeled anti-Ab added to well to bind to pt's Ab; excess washed away
• 4. colorless subst added to well; reacts w/ enz still there to produce a color

electrophoresis: separates mix of prot based on diff rates of migration in electric field (SDS-PAGE

pt antiserum incubated w/ blot, any Ab present that recog electrophoresed prot on blot bind; bound Ab can be detected w/ labeled anti-Ab to allow color rxns

sep a standardized prepartion of Ag derived from a particular pathogen and demonstrate "pattern" of Ab reactivity that provides a fingerprint unique to that pathogen

simultaneously demonstrate reactivities of mult Ab in polyclonal pt antiserum against mult Ag assoc w/ particular path

primary Ab (of known Ag specificity and labeled w/ fluorescein) reacts w/ specific Ag in test specimen

presence of an unlabeled "primary" Ab bound to Ag in a clinical specimen is detected by fluorescein-labeled "secondary" Ab; secondary finds and binds to primary

increased sensitivity of detection due to amplification effect as mult secondary Ab can bind to each single molecule of primary Ab

dep on bivalent characteristics of Ab that allows it to crosslink mol of Ag to each other; sol Ab bind to sol Ag molecules and lead to precip of lg insoluble Ag:Ab complexes...agglutination

agglutination test to detect reagin-type reactivity; uses carbon particles that are coated w/ cardiolipin Ag that reacts w/ reagin

Ab that react w/ lipid Ag assoc w/ Treponema

determines serum Ab titers in diagnosis of infectious diseases; demonstrates agglutination in tubes and prozone effect

conc of Ab is too high in relation to Ag (Ab interferes w/ its ability to crosslink); serum that is agglutination neg bc of prozone becomes pos when diluted

Ag:Ab rxn can lead to fixation of complement which triggers complement cascade and leads to cell lysis

complement binds to Ag:Ab complex; no lysis of RBC bc no free complement in serum bc all bound to Ag:Ab complex

no Ab present so no Ag:Ab complexes; free complement binds to RBC antibody and MAC forms

monolayers on culture plates exposed to pt specimens that may have virus; if virus in pt serum then it will infect cells in monolayer and make them lyse (CPE)...plaques form

viruses cannot infect monolayer bc of presence of virus-specfic Ab; no plaques; neutralizing Ab indicative of an Ab response against particular virus