ASSAYS

BY DON
IMMUNOASSAYS
USES ________
- SUBSTANCES THAT COMPLEX TO ANOTHER SUBSTANCE
- SUBSTANCE TO BE MEASURED

EXAMPLES:
-FLUORESCENCE
-RADIO
-ENZYME
FLUORESCENCE IMMUNOASSAY OR
___
ALBERT COONS 1941
FLUORESCENT PROBES USED:
GREEN FLUORESCENCE

RED ORANGE

RED

RED
DIRECT OR SINGLE LAYER (DFA)
IDENTIFIES _________
USES KNOWN ___________
POSITIVE RESULT:__________
INDIRECT OR DOUBLE LAYER
IDENTIFIES UNKNOWN ANTIBODY (DIRECT) OR UNKNOWN
ANTIGEN (INDIRECT)

USES KNOWN _____________

POSITIVE RESULT:__________
IMMUNOASSAYS
USES ________
- SUBSTANCES THAT COMPLEX TO ANOTHER SUBSTANCE
- SUBSTANCE TO BE MEASURED

EXAMPLES:
-FLUORESCENCE
-RADIO
-ENZYME
FLUORESCENCE IMMUNOASSAY OR
___
ALBERT COONS 1941
FLUORESCENT PROBES USED:
GREEN FLUORESCENCE

RED ORANGE

RED

RED
DIRECT OR SINGLE LAYER (DFA)
IDENTIFIES _________
USES KNOWN ___________
POSITIVE RESULT:__________
INDIRECT OR DOUBLE LAYER
IDENTIFIES UNKNOWN ANTIBODY (DIRECT) OR UNKNOWN
ANTIGEN (INDIRECT)

USES KNOWN _____________

POSITIVE RESULT:__________
FLUORESCENCE POLARIZATION
IA(FPIA)
-LABELED ANTIGENS COMPETE WITH UNLABELED ANTIGEN
IN THE PATIENT SAMPLE FOR A LIMITED NUMBER OF
ANTIBODY BINDING SITES
-DEGREE OF FLUORESCENCE POLARIZATION IS ____________
PROPORTIONAL TO CONCENTRATION OF THE ANALYTE
RADIOIMMUNOASSAY
-DEVELOPED BY ________ AND ________
-RADIOACTIVE ISOTOPES USED: _____,_______,_______,_____
-RADIOACTIVITY (RADIOACTIVE DECAY) IS MEASURED BY
______________
COMPETITIVE RIA
-ANALYTE BEING DETECTED COMPETES WITH A
RADIOLABELED ANALYTE FOR A LIMITED NUMBER OF
BINDING SITES

NOTE: AMOUNT OF LABEL IN BOUND PHASE IS

__________________________ TO THE AMOUNT OF PATIENT
ANTIGEN
-IF PATIENT HAS LITTLE AMOUNT OF ANTIGENS PRESENT, IT
WILL MAKE THE RADIOACTIVITY ________
-IF PATIENT HAS LOTS OF ANTIGENS PRESENT, IT WILL LEAD
TO ____________ RADIOACTIVITY
NONCOMPETITIVE RIA
-_____________ ASSAY
- ALSO CALLED IMMUNORADIOMETRIC ASSAY (IRMA)
NOTE: AMOUNT OF BOUND LABELED ANTIBODY IS
__________________TO THE AMOUNT OF PATIENT ANALYTE
RADIOIMMUNOASSAY TEST (RIST)
-FIRST DEVELOPED FOR THE MEASUREMENT OF ________
-COMPETITIVE RIA (OLDER METHOD)
-LARGELY BEEN REPLACED BY NONCOMPETITIVE SOLID-
PHASE IMMUNOASSAYS DUE TO THE EXPENSE AND
DIFFICULTY OF WORKING WITH RADIOACTIVITY
-IT SERVES AS A _______________ TEST TO DETERMINE IF
MORE SPECIFIC ALLERGY TESTING IS INDICATED
RADIOALLERGOSORBENT TEST(RAST)
-INTRODUCED IN ______
-NONCOMPETITIVE SOLID PHASE IMMUNOASSAY (OLDER
METHOD)
-PRINCIPLES OF THE TEST REMAIN THE SAME, BUT NEWER
TESTING METHODS INVOLVE THE USE OF ENZYME OR
FLUORESCENT LABELS RATHER THAN RADIOACTIVITY
ENZYME IMMUNOASSAY (EIA)
- INVOLVES ___________REACTION
RESULT: DARKER COLOR=_____________
ENZYME LABELS USED:

ALKALINE PHOSPHATASE YELLOW COLOR

HORSERADISH PEROXIDASE GREEN

GLUCOSE OXIDASE

BETA GALACTOSIDASE
ENZYME LINKED IMMUNO SORBENT
-RESULTS ARE READ USING ______________
-USED AS SCREENING METHOD FOR _____

INDIRECT ELISA= USES ___________

DOUBLE ANTIBODY ELISA= USES _________ AND

____________________
*FLUOROCHROMES ABSORB ENERGY FROM LIGHT SOURCE AND CONVERT IT TO LIGHT OF A
LONGER WAVELENGTH AND LOWER ENERGY
*CHEMILUMINESCENT MOLECULES PRODUCE LIGHT ENERGY FROM A CHEMICAL REACTION
RADIOISOTOPES ENZYMES FLUOROCHROMES CHEMILUMINESCENT
MOLECULES

125I

3H

14C