Department of Gastroenterology, Hepatology, Infectious Diseases and Endocrinology, Hannover Medical School, Carl-Neuberg-Straße 1, 30625, Hannover, Germany.
Member of the European Reference Network for Hepatological Diseases (ERN RARE-LIVER), Hamburg, Germany.
Sci Rep. 2023 Jun 20;13(1):10038. doi: 10.1038/s41598-023-37095-z.
Autoantibodies are the diagnostic hallmark of autoimmune liver diseases. Indirect immunofluorescence (IFT) is the reference method for the detection of anti-mitochondrial antibodies (AMA) and anti-liver kidney microsomal type-1 (anti-LKM1) antibodies, and inhibition ELISA (iELISA) for anti-soluble liver antigen (anti-SLA) antibodies. Given the complexity of these techniques, commercial ELISAs have emerged as a practical alternative, but without head-to-head validations. This study evaluated the agreement between three commercial ELISAs and the reference techniques and the impact of polyreactive immunoglobulin G (pIgG), a recently described phenomenon in autoimmune hepatitis, on commercial ELISAs. Inter-rater reliability was assessed using Cohen-Kappa coefficient (κ). Forty-eight, 46, and 66 samples were analyzed for AMA, anti-LKM1, and anti-SLA, respectively. For AMA, one commercial assay showed high agreement (κ = 0.91 (0.78-1.00)) with the reference method, while the other two showed weak or moderate agreement. For anti-LKM1, only one commercial assay showed high agreement (κ = 0.86 (0.71-1.0)). For anti-SLA antibodies only moderate agreement was achieved (κ up to 0.71 (0.52-0.89)). There was a trend towards higher pIgG levels in false-positives in the commercial ELISAs. Patients with high suspicion of autoimmune liver diseases should be referred to reference laboratories with the capacity of performing gold standard methods if the initial ELISA-based screening was performed.
自身抗体是自身免疫性肝病的诊断标志。间接免疫荧光(IFT)是检测抗线粒体抗体(AMA)和抗肝-肾微粒体 1 型(抗-LKM1)抗体的参考方法,抑制 ELISA(iELISA)用于检测抗可溶性肝抗原(抗-SLA)抗体。鉴于这些技术的复杂性,商业 ELISA 已成为一种实用的替代方法,但尚未进行头对头验证。本研究评估了三种商业 ELISA 与参考技术之间的一致性,以及最近在自身免疫性肝炎中描述的多反应性免疫球蛋白 G(pIgG)对商业 ELISA 的影响。采用 Cohen-Kappa 系数(κ)评估组内一致性。分别对 48、46 和 66 份 AMA、抗-LKM1 和抗-SLA 样本进行分析。对于 AMA,一种商业检测方法与参考方法具有高度一致性(κ=0.91(0.78-1.00)),而另外两种方法则显示出弱或中度一致性。对于抗-LKM1,只有一种商业检测方法显示出高度一致性(κ=0.86(0.71-1.0))。对于抗-SLA 抗体,仅达到中度一致性(κ 最高为 0.71(0.52-0.89))。在商业 ELISA 中的假阳性中,pIgG 水平呈上升趋势。如果最初进行了基于 ELISA 的筛选,而对自身免疫性肝病的高度怀疑,则应将患者转介至具有执行金标准方法能力的参考实验室。