Saliva is tested at IISBR in duplicate using a commercially-available enzymatic assay kit (ELISA) from Salimetrics following manufacturer’s protocol without modification. This test employs an un-treated 96-well microtitre plate. Uric Acid in standards, controls and unknown samples reacts with a proprietary reagent to produce a red chromogen over a 20 minute incubation. The resultant red color is read at 515nm on a spectrophotometer (PowerWave HT, BioTek Instruments). The amount of uric acid, as measured by the intensity of color, is directly proportional to the amount of uric acid present.
Briefly, 10 µL of standard, controls, and unknown saliva are pipetted in duplicate onto 96-well microtitre plates with 190ul uric acid reagent and incubate for 10 minutes shaking at 37⁰C. Then the plates incubate for 10 minutes at room temperature. The optical density (OD) read is at 515 nm. Concentrations of controls and samples are calculated from a standard proportion in which the kit manufacturer has established linearity from 0-20 mg/dL (Gen5, BioTek).