Table 1 Optimization conditions for oral fluid enzyme immunoassay

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