Description
Intended Uses: The Amphetamine Direct ELISA Kit provides only a preliminary analytical test result. A more specific alternate chemical method must be used in order to obtain a confirmed analytical result. Gas chromatography/ mass spectrometry (GS-MS) is the preferred confirmatory method (1). Professional judgment should be applied to any drug of abuse test result, particularly when preliminary positive results are used.
Principle of the Assay: The Amphetamine Direct ELISA Kit (for d-amphetamine measurement) is based upon the competitive binding to antibody of enzyme labeled antigen and unlabeled antigen, in proportion to their concentration in the reaction mixture. A 10 ul. aliquot of a diluted unknown specimen is incubated with a 100 ul. dilution of enzyme (Horseradish peroxidase) labeled d-amphetamine derivative in micro-plate wells, coated with fixed amounts of oriented high affinity purified polyclonal antibody. The wells are washed thoroughly and a chromogenic substrate added. The color produced is stopped using a dilute acid stop solution and the wells read at 450 nm. The intensity of the color developed is inversely proportional to the concentration of drug in the sample. The technique is sensitive to 1 ng/ml. The Amphetamine Direct ELISA Kit avoids extraction of urine sample for measurement. It employs a d-amphetamine directed antiserum. Due to the proprietary method of orienting the antibody on the polystyrene micro-plate much higher sensitivity is achieved compared to passive adsorption. This allows an extremely small sample size reducing matrix effects and interference with binding proteins(s) or other macromolecules