Table 1.

Optimization conditions for oral fluid enzyme immunoassay

Variable	Conditions Tested
Volume of solution in well	100 μL or 200 μL
Incubation time	1, 2, 6, 8, or 15 hours
Incubation temperature	4°C or 37°C
Dilution	OF alone, 1:2, 1:4 with PBST, 5% skim milk PBST
Proportion of sample on plate	75 μL OF + 25 μL diluent or 50 μL OF + 50 μL diluent
Centrifugation (5 min at 14000 rpm)	None, supernatant, resuspended pellet

OF = oral fluid

PBST = phosphate buffered saline with 0.05% Tween 20