MLT 269 Serology Study Guide Flashcards
Principle or clinical significance of direct fluorescent antibody test
A known Ab is labeled with a fluorescent dye and added to unknown Ag that is fixed to a slide. After incubation and was the slide is read using a fluorescent scope.
Ab-Ag rxn shows bright green fluorescence
List the principle and/or clinical significance of, in cluding reagents and pos and neg reactions for the ELISA
Competitive: enzyme labeled Ag competes w/ unlabeled pt Ag for binding sites on Ab molecules attached to a solid phase. The more pt Ag is bound the less enzyme labeled Ag can attach.
Non-competitive: higher sensitivity; measure status of immunity for certain agents and for autoantibody testing. Either Ag or Ab is bound to solid phase. When Ag is used for coating, pt Ab is added given time to react. After wash, enzyme labeled Ab is added. This 2nd Ab reacts w/ any pt Ab bound to solid phase. If no pt Ab is bound, the second Ab will not bind. After another wash, enzyme substrate is added. Color development is directly proportional to the amount of pt Ab.
List the principle and/or clinical significance of, in cluding reagents and pos and neg reactions for the complement fixation test
Complement is used as a reagent. Complement fixation occurs after the binding of Ag and Ab so complement uptake is an indicator of the presence of either specific Ag or Ab. Detects viral, fungal, or rickettsial Abs. Test uses an Ag and Ab, one of which is unknown, and sheep rbcs coated w/ hemolysin which will lyse in presence of complement. If lysis occurs in Stage 2, complement did not fix during stage 1.
List the principle and/or clinical significance of, in cluding reagents and pos and neg reactions for the RPR
Flocculation test for syphillis. Based on cardiolipin Ag technique but read macroscopically. Performed on unheated serum or plasma(does not require inactivation) w/ modified VDRL Ag and charcoal particles. Choline chloride replaces heat to inactivate complement.
List the principle and/or clinical significance of, in cluding reagents and pos and neg reactions for theFTA-ABS
patient serum absorbed with the extract of non-pathogenic Reiter treponeme to remove non-specific treponemal antibodies. a virulent strain of T pallidum(Ag) is allowed to react with the pt serum(Ab). specific T. pallidum Ab attached to the Nichols virulent strain of T. pallidum on a slide are demonstrated with a fluorescein-labeled antiglobulins (makes the Ag-Ab rxn visible) using dark field fluorescence microscopy.
List the principle and/or clinical significance of, in cluding reagents and pos and neg reactions for the differential absorption test of Davidson
determination of anti-sheep antibodies after absorption with guinea pig kidney and possibly beef RBCs. A positive reaction is indicated by complete removal of anti-sheep agglutinins(Ab) by guinea pig kidney and/or beef RBC/s
List the principle and/or clinical significance of, in cluding reagents and pos and neg reactions for the RA test
SCREEN: Rheumatoid factor (RF) Ab is found in 80% of pts. Uses agglutination test with latex particles coated with IgG.
DIAGNOSTIC: EIA technique or Nephelometric assays used to test for IgG or IgA isotypes which rarely occur in other diseases. Disease activity is followed by measuring inflammation thru ESR, CRP and complement components.
List the principle and/or clinical significance of, in cluding reagents and pos and neg reactions for the ANA test.
SCREEN: fluorescent Antinuclear AB; mouse kidney or human epithelial HEp-2 cells are fixed to a slide and allowed to react with pt serum. Fluorescent tagged antihuman immunoglobulin or horseradish peroxidase is added(conjugate)
DIAGNOSTIC: Double stranded DNA Ab are seen only in lupus(50-70%) and levels correlate with disease activity. Produces peripheral or homogenous staining pattern on indirect immunofluorescence.
List the principle and/or clinical significance of, in cluding reagents and pos and neg reactions for cold agglutinins
Keep blood sample at 37 deg C until serum and cells separate. Make serial dilutions of sample serum with human group O as the Ag. Incubate overnight in fridge and view for agglutination. Warm to 37 deg C for 30 mins If agg disappears, then cold agg are present..
High titers that do not vary over time may indidcate mycoplasma pneumonia, IM, autoimmune hemolytic anemia, and viral infections(mumps, rubella, flu, HIV…)
What is the causative agent of syphillis?
IM?
Treponemal pallidum
Epstein-Barr virus
Ig produced first after exposure to an antigen
IgM
Ig found in greatest concentration
IgG
Ig appearing in greatest concentration to a second antigen exposure
IgG
Ig associated with allergic reations
IgE
List the differential absorption characteristics of IM(non-Forssman), Forssman, and serum sickness antibodies.
HAb GPkidney Beef rbc
Forss Yes No
IM No Yes
SeSi Yes Yes
reagin
Ab found in blood of a person with a genetic predisposition for allergies; hay fever; IgE response
Ab formed against lipid material from damaged cells; substance found in the blood of an individual that has a positive reaction to serological syphillis test.
anamnestic reaction
secondary immune response; IgG; 1-2 weeks after primary exposure, faster response, stays higher, longer
Prozone
the failure of a serological reaction to occur, due to antiBody excess