Budelier Melissa M, Franks Caroline E, Logsdon Nicole, Jannetto Paul J, Scott Mitchell G, Roper Stephen M, Farnsworth Christopher W
Department of Pathology and Immunology, Division of Laboratory and Genomic Medicine, Washington University, St. Louis, MO.
Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN.
J Appl Lab Med. 2020 Nov 1;5(6):1277-1286. doi: 10.1093/jalm/jfaa048.
Fentanyl is a synthetic opioid associated with illicit drug use and overdose deaths. The SEFRIA Immunalysis (IAL) and ARK fentanyl assays are both FDA-cleared, open channel immunoassays for fentanyl detection in urine. However, limited data are available in the literature comparing these assays. The objective of this study was to perform a direct comparison of these two fentanyl immunoassays.
IAL and ARK fentanyl immunoassays were performed on a Roche Cobas e602 automated chemistry analyzer. Repeatability and total imprecision were compared by diluting fentanyl into urine at concentrations above, below, and at the manufacturers' cutoffs of 1.0 ng/mL. Cross-reactivity was assessed for norfentanyl and the fentanyl analogs acetylfentanyl, acrylfentanyl, and furanylfentanyl. Concordance was assessed in 90 patient samples using liquid chromatography-tandem mass spectrometry (LC-MS/MS) as the gold standard.
Repeatability varied from 11.4%-17.8% on the IAL assay and 2.8%-5.5% on the ARK assay. Total imprecision was 18.9%-40.7% on the IAL assay and 2.9%-6.4% on the ARK assay. Both assays cross-reacted with acetylfentanyl (∼100%), acrylfentanyl (∼100%), and furanylfentanyl (∼20%), but only the ARK assay cross-reacted with norfentanyl (∼3%). An admixture of 0.5 ng/mL fentanyl and 6 ng/mL norfentanyl produced a positive result on the ARK assay. Total concordance between IAL and ARK for 90 tested patient samples was 93% (kappa = 0.85). Relative to LC-MS/MS, the IAL assay had a concordance of 90% (kappa = 0.79) and the ARK assay had a concordance of 94% (kappa = 0.88). Including norfentanyl in the LC-MS/MS confirmation increased the concordance of the ARK to 96% (kappa = 0.90).
The ARK assay recognized the metabolite norfentanyl, demonstrated superior precision, and had better concordance with LC-MS/MS compared to the IAL assay.
芬太尼是一种与非法药物使用和过量死亡相关的合成阿片类药物。SEFRIA免疫分析（IAL）和ARK芬太尼检测法均为经美国食品药品监督管理局（FDA）批准的用于尿液中芬太尼检测的开放通道免疫分析法。然而，文献中比较这两种检测法的数据有限。本研究的目的是对这两种芬太尼免疫分析法进行直接比较。
IAL和ARK芬太尼免疫分析法在罗氏Cobas e602自动化学分析仪上进行。通过将芬太尼稀释到高于、低于制造商设定的1.0 ng/mL临界值以及临界值浓度的尿液中，比较重复性和总不精密度。评估去甲芬太尼以及芬太尼类似物乙酰芬太尼、丙烯酰芬太尼和呋喃芬太尼的交叉反应性。使用液相色谱 - 串联质谱法（LC-MS/MS）作为金标准，对90份患者样本评估一致性。
IAL检测法的重复性在11.4% - 17.8%之间，ARK检测法的重复性在2.8% - 5.5%之间。IAL检测法的总不精密度为18.9% - 40.7%，ARK检测法的总不精密度为2.9% - 6.4%。两种检测法均与乙酰芬太尼（约100%）、丙烯酰芬太尼（约100%）和呋喃芬太尼（约20%）发生交叉反应，但只有ARK检测法与去甲芬太尼（约3%）发生交叉反应。0.5 ng/mL芬太尼和6 ng/mL去甲芬太尼的混合物在ARK检测法中产生阳性结果。90份受试患者样本中IAL和ARK的总一致性为93%（kappa = 0.85）。相对于LC-MS/MS，IAL检测法的一致性为90%（kappa = 0.79），ARK检测法的一致性为94%（kappa = 0.88）。在LC-MS/MS确认中纳入去甲芬太尼后，ARK的一致性提高到96%（kappa = 0.90）。
与IAL检测法相比，ARK检测法能识别代谢物去甲芬太尼，精密度更高，与LC-MS/MS的一致性更好。
