Evaluation of enzyme-linked immunosorbent assay screening kits for the detection of nitazene analogs.

Amanda L Pacana, Britni N Skillman
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Abstract

Nitazene analogs are a highly potent class of novel psychoactive substances (NPS) that were first identified in forensic casework in the United States in 2019. While enzyme-linked immunosorbent assay (ELISA) remains a prevalent screening tool in forensic toxicology laboratories, no nitazene-specific ELISA kits are commercially available, supporting the use of high-resolution mass spectrometry (HRMS) methods as a more adaptable alternative for screening. However, even with the growth in popularity of HRMS techniques, it is important to understand the cross-reactivity of novel substances, such as nitazenes, with routinely used ELISA kits. Cross-reactivity is particularly important for forensic toxicology casework since it can impact the reliability of screening results, potentially leading to non-detection of novel substances or false-positive identifications in the presence of non-target analytes. This study tested the cross-reactivity of seven nitazene analogs (4'-OH nitazene, 5-methyl etodesnitazene, isotonitazene, metodesnitazene, N-piperidinyl etonitazene, N-pyrrolidino etonitazene, and protonitazene) in whole blood using 13 commercial ELISA kits from three manufacturers. Various drug/class kits were selected based on reported or potential co-positivity with nitazenes (opiates, opioids, fentanyl) or by structural similarities (LSD, zolpidem). Across tested concentrations for the seven selected analytes, none of the tested kits produced a signal sufficient for a positive result, confirming that their presence at these levels does not compromise the screening specificity of the target analytes. However, these findings highlight the need for laboratories to adopt mass spectral-based screening methods like HRMS or advocate for the development of nitazene-specific ELISA kits for effective nitazene analog screening.

酶联免疫吸附法检测nitazene类似物筛选试剂盒的评价。
Nitazene类似物是一类高效的新型精神活性物质(NPS),于2019年在美国的法医案件中首次被发现。虽然酶联免疫吸附测定法(ELISA)仍然是法医毒理学实验室中普遍使用的筛查工具,但市面上没有nitazen特异性ELISA试剂盒,这支持使用高分辨率质谱法(HRMS)作为更适用的筛查替代方法。然而,即使随着HRMS技术的普及,了解新物质(如nitazene)与常规ELISA试剂盒的交叉反应性也很重要。交叉反应性对于法医毒理学案例工作尤其重要,因为它会影响筛选结果的可靠性,可能导致无法检测到新物质或在存在非目标分析物的情况下产生假阳性鉴定。本研究使用三家厂商生产的13套商用ELISA试剂盒,检测了7种硝基类似物(4'-OH硝基、5-甲基乙硝基、异硝基、甲硝基、n -哌啶基乙硝基和原硝基)在全血中的交叉反应性。根据与nitazenes(阿片类药物、阿片类药物、芬太尼)报告的或潜在的共阳性或结构相似性(LSD、唑吡坦)选择各种药物/类别试剂盒。在七种选定分析物的测试浓度中,没有一种测试试剂盒产生足以产生阳性结果的信号,证实它们在这些水平下的存在不会损害目标分析物的筛选特异性。然而,这些发现强调了实验室采用质谱筛选方法(如HRMS)或倡导开发nitazene特异性ELISA试剂盒以有效筛选nitazene类似物的必要性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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