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Validation of an ELISA method for screening methadone in postmortem blood.
J Anal Toxicol 2006; 30(8):617-20JA

Abstract

In this study, we evaluate Venture Labs' enzyme-linked immunosorbent assay (ELISA) for the detection of methadone in postmortem specimens. Sixty-one postmortem specimens that previously screened positive for methadone along with 59 specimens which screened negative for methadone were included. All specimens were screened using the Venture Labs methadone assay in conjunction with a liquid-liquid basic extraction and gas chromatographic-mass spectrometric (GC-MS) analysis. All cases screening positive by either method were confirmed for methadone and its metabolite 2-ethylidene-1,5-dimethyl- 3,3-diphenylpyrrolidine by a solid-phase extraction utilizing deuterated internal standards and GC-MS-SIM. Twenty-four postmortem samples that screened negative by both methods were also extracted and analyzed using the confirmation method to demonstrate the validity of both screening methods. The intra- and interassay precision for the ELISA method was evaluated at the cut-off concentration used for the analysis (50 ng/mL). True positives, true negatives, false positives, and false negatives were calculated for the ELISA results as compared to the GC-MS screening data. The Venture Labs methadone assay demonstrated a sensitivity of 96.7%+/-2.3% and a specificity of 98.3%+/-1.7% relative to the GC-MS method.

Authors+Show Affiliations

UMass Memorial Forensic Toxicology Laboratory, Worcester, Massachusetts 01605, USA.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Validation Studies

Language

eng

PubMed ID

17132262

Citation

Juhascik, Matthew, et al. "Validation of an ELISA Method for Screening Methadone in Postmortem Blood." Journal of Analytical Toxicology, vol. 30, no. 8, 2006, pp. 617-20.
Juhascik M, Habbel S, Barron W, et al. Validation of an ELISA method for screening methadone in postmortem blood. J Anal Toxicol. 2006;30(8):617-20.
Juhascik, M., Habbel, S., Barron, W., & Behonick, G. (2006). Validation of an ELISA method for screening methadone in postmortem blood. Journal of Analytical Toxicology, 30(8), pp. 617-20.
Juhascik M, et al. Validation of an ELISA Method for Screening Methadone in Postmortem Blood. J Anal Toxicol. 2006;30(8):617-20. PubMed PMID: 17132262.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Validation of an ELISA method for screening methadone in postmortem blood. AU - Juhascik,Matthew, AU - Habbel,Steven, AU - Barron,William, AU - Behonick,George, PY - 2006/11/30/pubmed PY - 2006/12/15/medline PY - 2006/11/30/entrez SP - 617 EP - 20 JF - Journal of analytical toxicology JO - J Anal Toxicol VL - 30 IS - 8 N2 - In this study, we evaluate Venture Labs' enzyme-linked immunosorbent assay (ELISA) for the detection of methadone in postmortem specimens. Sixty-one postmortem specimens that previously screened positive for methadone along with 59 specimens which screened negative for methadone were included. All specimens were screened using the Venture Labs methadone assay in conjunction with a liquid-liquid basic extraction and gas chromatographic-mass spectrometric (GC-MS) analysis. All cases screening positive by either method were confirmed for methadone and its metabolite 2-ethylidene-1,5-dimethyl- 3,3-diphenylpyrrolidine by a solid-phase extraction utilizing deuterated internal standards and GC-MS-SIM. Twenty-four postmortem samples that screened negative by both methods were also extracted and analyzed using the confirmation method to demonstrate the validity of both screening methods. The intra- and interassay precision for the ELISA method was evaluated at the cut-off concentration used for the analysis (50 ng/mL). True positives, true negatives, false positives, and false negatives were calculated for the ELISA results as compared to the GC-MS screening data. The Venture Labs methadone assay demonstrated a sensitivity of 96.7%+/-2.3% and a specificity of 98.3%+/-1.7% relative to the GC-MS method. SN - 0146-4760 UR - https://www.unboundmedicine.com/medline/citation/17132262/Validation_of_an_ELISA_method_for_screening_methadone_in_postmortem_blood_ L2 - https://academic.oup.com/jat/article-lookup/doi/10.1093/jat/30.8.617 DB - PRIME DP - Unbound Medicine ER -