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Validation of an ELISA-based screening assay for the detection of amphetamine, MDMA and MDA in blood and oral fluid.
Forensic Sci Int 2005; 153(1):29-37FS

Abstract

The use of amphetamine and 'ecstasy' (MDMA) has increased exponentially in many European countries since the late nineties, leading to a rapid growth in the number of clinical and forensic analyses. Therefore, a rapid screening procedure for these substances in biological specimens has become an important part of routine toxicological analysis in forensic laboratories. The objective of this study was to evaluate the Cozart amphetamine enzyme-linked immunosorbent assay (ELISA) for the screening of plasma samples and oral fluid samples (collected with the Intercept device). Authentic plasma samples from drivers (n=360) were screened, using an 1:5-fold dilution. True positive, true negative, false positive and false negative results were determined relative to the in-house routine GC-MS analysis. Samples consisted of 144 amphetamine-only positives, 141MDMA/MDA-only positives, and 74 negatives when using the limit of quantitation as the cut-off level for confirmation (10 ng/mL). Using these results, receiver operating characteristic (ROC) curves were generated and optimal cut-off values for the screening assay were calculated. Analysis showed that the ELISA is able to predict the presence of either amphetamine or *MDMA/MDA (*MDMA as its metabolite MDA) in plasma samples with 98.3% sensitivity and 100% specificity at a cut-off value of 66.5 ng/mL d-amphetamine equivalents. A similar analysis was conducted on 216 oral fluid specimens collected from a controlled double blind study. Subjects received placebo or a high (100 mg) or low (75 mg) dose of MDMA. Oral fluid samples were collected at 1.5 and 5.5h after administration. Combined results of the analysis of the high and low dose oral fluid samples indicated a screening cut-off of 51 ng/mL d-amphetamine equivalents with both a sensitivity and specificity of 98.6% (using a LC-MS/MS confirmation cut-off of 10 ng/mL). In conclusion, these data indicate that the Cozart AMP EIA plates constitute a fast and accurate screening technique for the identification of amphetamine and MDMA/MDA positive plasma samples and oral fluid specimens (collected with Intercept. It should be emphasized that method validation should be performed for each type of biological matrix.

Authors+Show Affiliations

Federal Public Service Justice, National Institute of Criminalistics and Criminology (NICC), Section Toxicology, Vilvoordsesteenweg 98, 1120 Brussels, Belgium. marleen.laloup@just.fgov.beNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Clinical Trial
Controlled Clinical Trial
Journal Article
Validation Studies

Language

eng

PubMed ID

15922530

Citation

Laloup, Marleen, et al. "Validation of an ELISA-based Screening Assay for the Detection of Amphetamine, MDMA and MDA in Blood and Oral Fluid." Forensic Science International, vol. 153, no. 1, 2005, pp. 29-37.
Laloup M, Tilman G, Maes V, et al. Validation of an ELISA-based screening assay for the detection of amphetamine, MDMA and MDA in blood and oral fluid. Forensic Sci Int. 2005;153(1):29-37.
Laloup, M., Tilman, G., Maes, V., De Boeck, G., Wallemacq, P., Ramaekers, J., & Samyn, N. (2005). Validation of an ELISA-based screening assay for the detection of amphetamine, MDMA and MDA in blood and oral fluid. Forensic Science International, 153(1), pp. 29-37.
Laloup M, et al. Validation of an ELISA-based Screening Assay for the Detection of Amphetamine, MDMA and MDA in Blood and Oral Fluid. Forensic Sci Int. 2005 Oct 4;153(1):29-37. PubMed PMID: 15922530.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Validation of an ELISA-based screening assay for the detection of amphetamine, MDMA and MDA in blood and oral fluid. AU - Laloup,Marleen, AU - Tilman,Gaëlle, AU - Maes,Viviane, AU - De Boeck,Gert, AU - Wallemacq,Pierre, AU - Ramaekers,Jan, AU - Samyn,Nele, PY - 2005/6/1/pubmed PY - 2005/9/24/medline PY - 2005/6/1/entrez SP - 29 EP - 37 JF - Forensic science international JO - Forensic Sci. Int. VL - 153 IS - 1 N2 - The use of amphetamine and 'ecstasy' (MDMA) has increased exponentially in many European countries since the late nineties, leading to a rapid growth in the number of clinical and forensic analyses. Therefore, a rapid screening procedure for these substances in biological specimens has become an important part of routine toxicological analysis in forensic laboratories. The objective of this study was to evaluate the Cozart amphetamine enzyme-linked immunosorbent assay (ELISA) for the screening of plasma samples and oral fluid samples (collected with the Intercept device). Authentic plasma samples from drivers (n=360) were screened, using an 1:5-fold dilution. True positive, true negative, false positive and false negative results were determined relative to the in-house routine GC-MS analysis. Samples consisted of 144 amphetamine-only positives, 141MDMA/MDA-only positives, and 74 negatives when using the limit of quantitation as the cut-off level for confirmation (10 ng/mL). Using these results, receiver operating characteristic (ROC) curves were generated and optimal cut-off values for the screening assay were calculated. Analysis showed that the ELISA is able to predict the presence of either amphetamine or *MDMA/MDA (*MDMA as its metabolite MDA) in plasma samples with 98.3% sensitivity and 100% specificity at a cut-off value of 66.5 ng/mL d-amphetamine equivalents. A similar analysis was conducted on 216 oral fluid specimens collected from a controlled double blind study. Subjects received placebo or a high (100 mg) or low (75 mg) dose of MDMA. Oral fluid samples were collected at 1.5 and 5.5h after administration. Combined results of the analysis of the high and low dose oral fluid samples indicated a screening cut-off of 51 ng/mL d-amphetamine equivalents with both a sensitivity and specificity of 98.6% (using a LC-MS/MS confirmation cut-off of 10 ng/mL). In conclusion, these data indicate that the Cozart AMP EIA plates constitute a fast and accurate screening technique for the identification of amphetamine and MDMA/MDA positive plasma samples and oral fluid specimens (collected with Intercept. It should be emphasized that method validation should be performed for each type of biological matrix. SN - 0379-0738 UR - https://www.unboundmedicine.com/medline/citation/15922530/Validation_of_an_ELISA_based_screening_assay_for_the_detection_of_amphetamine_MDMA_and_MDA_in_blood_and_oral_fluid_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0379-0738(05)00209-4 DB - PRIME DP - Unbound Medicine ER -