| Development of a homogeneous immunoassay for the detection of fentanyl in urine. | |
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MedLine Citation:
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PMID: 20801588 Owner: NLM Status: In-Data-Review |
Abstract/OtherAbstract:
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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 "in-house" 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. |
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Authors:
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Guohong Wang; Kim Huynh; Rekha Barhate; Warren Rodrigues; Christine Moore; Cynthia Coulter; Michael Vincent; James Soares |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Forensic science international Volume: 206 ISSN: 1872-6283 ISO Abbreviation: Forensic Sci. Int. Publication Date: 2011 Mar |
Date Detail:
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Created Date: 2011-03-14 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 7902034 Medline TA: Forensic Sci Int Country: Ireland |
Other Details:
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Languages: eng Pagination: 127-31 Citation Subset: IM |
Copyright Information:
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Copyright © 2010 Elsevier Ireland Ltd. All rights reserved. |
Affiliation:
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Immunalysis Corporation, 829 Towne Center Drive, Pomona, CA 91767, USA. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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