Description
INTENDED USE
The Calbiotech, Inc. (CBI) Canine Fentanyl Direct ELISA Kit is a sensitive in-vitro test to detect the presence of Fentanyl in Canine serum or urine.
SUMMARY AND EXPLANATION
Fentanyl is a synthetic narcotic analgesic of high potency and short duration of action. Though 200 times more potent than morphine, Fentanyl has a high safety margin. The drug is available as a citrate salt in an injectable solution containing 50 μg/ml. It is also available as a transdermal patch containing 2.5 – 10 mg Fentanyl and provides a dose of 25 –100 μg/hr for 72 hours for management of chronic pain.(1) While Fentanyl has all the properties of morphine, it is structurally different and therefore cannot be detected by screening tests for morphine and related opiates. Because of the potency of the drug, concentrations encountered in biological fluids are in the sub nanogram range.
PRINCIPLE OF THE TEST
The Canine Fentanyl Direct ELISA Kit 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 20 μl. aliquot of a diluted unknown specimen is incubated with a 100 μl. dilution of enzyme (Horseradish peroxidase) labeled Fentanyl derivative in micro-plate wells, coated with fixed amounts of high affinity purified polyclonal anti-
Fentanyl. 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 0.1 ng/ml. The Canine Fentanyl Direct ELISA Kit avoids extraction of urine or blood sample for measurement. It employs a Fentanyl 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.
