Automated immunohistochemistry of intra-epidermal nerve fibres in skin biopsies: A proof-of-concept study

IF 3.9 3区医学 Q1 CLINICAL NEUROLOGY Journal of the Peripheral Nervous System Pub Date : 2024-08-20 DOI:10.1111/jns.12650

Jamie Burgess, Anne Marshall, Leandros Rapteas, J. Hamill Kevin, Andrew Marshall, Rayaz A. Malik, Bernhard Frank, Uazman Alam

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Abstract

Aims

To develop a standardised, automated protocol for detecting protein gene product 9.5 (PGP9.5) positive intra-epidermal nerve fibres (IENFs) in skin biopsies, transitioning from the established manual technique to an automated platform. This automated method, although currently intended for research applications, may improve the accessibility of this diagnostic test for small fibre neuropathy in clinical settings.

Methods

Skin biopsies (n = 274) from 100 participants (fibromyalgia syndrome n = 62; idiopathic small fibre neuropathy: n = 16; healthy volunteers: n = 22) were processed using an automated immunohistochemistry platform. IENF quantification was performed by blinded examiners, with reliability assessed via a two-way mixed-effects model to evaluate inter- and intra-observer variability.

Results

The automated staining system reproduced intra-epidermal nerve fibre density (IENFD) counts consistent with free-floating sections (mean ± standard deviation: free-floating: 5.6 ± 3.4 fibres/mm; automated: 5.9 ± 3.2 fibres/mm). A median difference of 0.3 with a lower bound 95% Confidence Interval (CI) at −0.00005 established non-inferiority against a margin of −0.4 (p = .08). Specifically, the inter-class correlation coefficient (class denotes consistency in measured observations) was 99% (95% CI: 0.9–1), indicating excellent agreement between free-floating and automated methods. The inter- and intra-class coefficient between examiners were both 99% (95% CI: 0.9–0.1) for IENFD, demonstrating high reliability using sections stained using the automated method.

Interpretation

Automated immunohistochemistry provides high-throughput reliable and reproducible intra-epidermal nerve fibre quantification. This method, although currently proof-of-concept, for research use only, may be more widely deployed in histopathology laboratories to increase the adoption of IENFD assessment for the diagnosis of peripheral neuropathies.

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皮肤活检中表皮内神经纤维的自动免疫组化：概念验证研究

目的：开发一种用于检测皮肤活组织切片中表皮内神经纤维（IENFs）蛋白基因产物 9.5（PGP9.5）阳性的标准化自动化方案，将既有的手工技术过渡到自动化平台。尽管这种自动化方法目前主要用于研究应用，但它可能会提高这种小纤维神经病诊断检测在临床环境中的可及性：方法：使用自动免疫组化平台处理 100 名参与者（纤维肌痛综合征：62 人；特发性小纤维神经病：16 人；健康志愿者：22 人）的皮肤活检组织（n = 274）。IENF 定量由双盲检验员进行，可靠性通过双向混合效应模型进行评估，以评价观察者之间和观察者内部的变异性：结果：自动染色系统再现的表皮内神经纤维密度（IENFD）计数与自由浮动切片一致（平均值±标准偏差：自由浮动切片：5.6±3.4 纤维）：5.6 ± 3.4 纤维/毫米；自动：5.9 ± 3.2 纤维/毫米：5.9 ± 3.2 纤维/毫米）。中位数差异为 0.3，95% 置信区间 (CI) 下限为 -0.00005，与 -0.4 的差值相比，确定了非劣效性（p = .08）。具体来说，类间相关系数（类表示测量观察结果的一致性）为 99%（95% 置信区间：0.9-1），表明自由浮动法和自动化方法之间的一致性极佳。对于 IENFD，检查者之间的类间和类内系数均为 99%（95% CI：0.9-0.1），这表明使用自动方法染色的切片具有很高的可靠性：自动免疫组化可提供高通量、可靠且可重复的表皮内神经纤维定量。这种方法虽然目前只是概念验证，仅供研究使用，但可以更广泛地应用于组织病理学实验室，以提高周围神经病诊断中 IENFD 评估的采用率。

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来源期刊

Journal of the Peripheral Nervous System 医学-临床神经学

CiteScore

6.10

自引率

7.90%

发文量

审稿时长

>12 weeks

期刊介绍： The Journal of the Peripheral Nervous System is the official journal of the Peripheral Nerve Society. Founded in 1996, it is the scientific journal of choice for clinicians, clinical scientists and basic neuroscientists interested in all aspects of biology and clinical research of peripheral nervous system disorders. The Journal of the Peripheral Nervous System is a peer-reviewed journal that publishes high quality articles on cell and molecular biology, genomics, neuropathic pain, clinical research, trials, and unique case reports on inherited and acquired peripheral neuropathies. Original articles are organized according to the topic in one of four specific areas: Mechanisms of Disease, Genetics, Clinical Research, and Clinical Trials. The journal also publishes regular review papers on hot topics and Special Issues on basic, clinical, or assembled research in the field of peripheral nervous system disorders. Authors interested in contributing a review-type article or a Special Issue should contact the Editorial Office to discuss the scope of the proposed article with the Editor-in-Chief.