@article{ref1,
title="Development of a homogeneous immunoassay for the detection of fentanyl in urine",
journal="Forensic science international",
year="2011",
author="Wang, Guohong and Huynh, Kim and Barhate, Rekha and Rodrigues, Warren and Moore, Christine and Coulter, Cynthia and Vincent, Michael and Soares, James",
volume="206",
number="1-3",
pages="127-131",
abstract="OBJECTIVE: Fentanyl is an extremely potent synthetic opioid that is widely used for chronic pain treatment; it is highly addictive and prone to abuse. The objective is to develop a high throughput homogeneous enzyme immunoassay (HEIA) for the rapid detection of fentanyl in human urine. METHODS: The HEIA is based on an immunoassay format in which both the antibody and enzyme-drug conjugate are in ready-to-use solution. In the absence of the target analyte in the specimen, enzyme-labeled drug conjugate binds to the antibody and results in a decrease of the enzyme (G6PDH) activity; hence there is lower absorbance at 340nm. If the target analyte is present in the specimen, it competes with the enzyme-labeled drug to bind to limited amount of specific antibody that result in more enzyme activity and yields an increased absorbance at 340nm. A polyclonal &quot;in-house&quot; antibody was selected that is capable of measuring fentanyl at low concentrations thus the assay detection limit was determined to be 1ng/mL. The assay was validated with clinical urine specimens that previously confirmed positively or negatively for fentanyl/norfentanyl by LC-MS/MS. RESULTS: The intra-day (n=20) and inter-day (n=100) precision of the assay was less than 1% CV. No interferences from structurally unrelated and commonly ingested drugs were observed at a concentration of 10,000ng/mL. A total of 209 LC-MS/MS confirmed urine specimens (149 positive and 57 negative samples) were analyzed by HEIA. The sensitivity, specificity, and accuracy values were 99%, 95%, and 98% respectively. CONCLUSION: This paper describes the development of a highly sensitive homogenous enzyme immunoassay for detecting fentanyl in urine at a cut-off concentration of 2ng/mL.<p /> <p>Language: en</p>",
language="en",
issn="0379-0738",
doi="10.1016/j.forsciint.2010.07.022",
url="http://dx.doi.org/10.1016/j.forsciint.2010.07.022"
}