King's Forensics, Department of Analytical, Environmental & Forensic Sciences, Franklin-Wilkins Building, King's College London, 150 Stamford St, London, United Kingdom, SE1 9NH, UK.
Intelligent Fingerprinting, Cambridge, UK.
Drug Test Anal. 2022 Apr;14(4):613-621. doi: 10.1002/dta.3196. Epub 2021 Dec 8.
To date, a specific point-of-care test (POCT) for 3,4-methylenedioxymethamphetamine (MDMA, ecstasy, 'E') in latent fingerprints (LFPs) has not been explored. Other POCTs identify MDMA in sweat by detecting the drug as a cross-reactant rather than target analyte, thus decreasing the test's sensitivity. The study's aim was to design a sensitive POCT for the detection of MDMA in LFPs using surface plasmon resonance (SPR) and lateral flow immunoassay (LFA) technology. A high-affinity antibody binding pair was identified using the former technique, deeming the pair suitable for a LFA. Titrations of fluorescently labelled antibody and antigen concentrations were tested to identify a sharp drop-in signal upon the addition of MDMA to allow a clear distinction between negative and positive outcomes. We trialled the LFA by producing dose response curves with MDMA and a group of drugs that share a similar chemical structure to MDMA. These were generated through spiking the LFA with increasing levels of drug (0-400 pg/10 μl of MDMA; 0-10,000 pg/10 μl of cross-reactant). Fluorescent test signals were measured using a cartridge reader. The cut-off (threshold) 60 pg/10 μl calculated better cartridge performance (1.00 sensitivity, 0.95 specificity and 0.98 accuracy), when compared with 40 pg/10 μl. The biggest cross-reactant was PMMA (250%), followed by MDEA (183%), MBDB (167%), MDA (16%) and methamphetamine (16%). A sensitive LFP screening tool requiring no sample preparation was successfully designed.
迄今为止,尚未探索用于潜伏指纹(LFPs)中 3,4-亚甲二氧基甲基苯丙胺(MDMA,摇头丸,“E”)的特定即时检验(POCT)。其他 POCT 通过检测药物作为交叉反应物而不是目标分析物来识别汗水中的 MDMA,从而降低了测试的灵敏度。本研究旨在设计一种使用表面等离子体共振(SPR)和侧向流动免疫测定(LFA)技术灵敏检测 LFPs 中 MDMA 的 POCT。使用前者技术鉴定出高亲和力的抗体结合对,认为该对适合 LFA。测试荧光标记的抗体和抗原浓度的滴定,以鉴定在添加 MDMA 时信号明显下降,从而可以清楚地区分阴性和阳性结果。我们通过用 MDMA 和一组具有相似化学结构的药物对 LFA 进行加标,来测试 LFA。这些是通过用 LFA 逐渐增加药物水平(0-400 pg/10 μl 的 MDMA;0-10,000 pg/10 μl 的交叉反应物)产生的。荧光测试信号使用盒式阅读器进行测量。与 40 pg/10 μl 相比,计算出的 60 pg/10 μl (阈值)更能改善盒式性能(1.00 灵敏度,0.95 特异性和 0.98 准确性)。最大的交叉反应物是 PMMA(250%),其次是 MDEA(183%),MBDB(167%),MDA(16%)和甲基苯丙胺(16%)。成功设计了一种无需样品制备的灵敏 LFPs 筛选工具。