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A novel immunoassay for quantitative drug abuse screening in serum

2016; Elsevier BV; Volume: 436; Linguagem: Inglês

10.1016/j.jim.2016.06.004

1872-7905

Sarah Schumacher, Harald Seitz,

Monoclonal and Polyclonal Antibodies Research

An immunoassay was established which enables a reliable quantification of serological drug samples. The assay is based on a competitive ELISA. In total nine drugs (amphetamine, methamphetamine, 3,4-methylenedioxy-methamphetamine (MDMA), tetrahydrocannabinol (THC), phencyclidine (PCP), methadone, morphine, cocaine and benzoylecgonine) were tested. All reagents had to pass through a stringent validation process. Within the established test for three out of the nine drugs no cross-reactivity with any tested compounds, e.g. serum, other antibodies or chemically related molecules was detectable for the tested antibodies. Furthermore, a sensitive and selective detection was possible, even in the presence of up to 9 drugs or of various anti-drug antibodies. After exclusion of cross-reactivities antibodies against three drugs (methadone, MDMA, benzoylecgonine) were validated, which allowed a specific and sensitive quantification. For the competitive measurements CVs in the range of 2–17% could be reached with LLOQs of 10 ng/mL and LODs of 150 ng/mL for methadone, 250 ng/mL for MDMA and 400 ng/mL for benzoylecgonine. Anonymized serum samples (n = 10) provided by the office of criminal investigation Berlin were analyzed for verification purposes. Evaluation of these data showed a correlation (CV) of ≈ 0.9 with standard GC–MS methods. A miniaturization on microarray was possible by using the anti-MDMA antibody for the detection of MDMA in serum. The microarray increased the through-put drastically and enabled the simultaneous quantification of various drugs.