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

Burgess, Jamie ORCID: 0000-0002-7165-6918, Marshall, Anne ORCID: 0000-0002-2331-3750, Rapteas, Leandros, Kevin, J Hamill, Marshall, Andrew ORCID: 0000-0001-8273-7089, Malik, Rayaz A, Frank, Bernhard ORCID: 0000-0001-5405-4942 and Alam, Uazman ORCID: 0000-0002-3190-1122 (2024) Automated immunohistochemistry of intra‐epidermal nerve fibres in skin biopsies: A proof‐of‐concept study Journal of the Peripheral Nervous System, 29 (3). pp. 329-338. ISSN 1085-9489, 1529-8027

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Abstract

AbstractAimsTo 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.MethodsSkin 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.ResultsThe 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.InterpretationAutomated 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.

Item Type:	Article
Uncontrolled Keywords:	automated, density, digital pathology, fibromyalgia syndrome, immunohistochemistry, intra-epidermal nerve fibres, pathology, quantification, small fibre neuropathy
Depositing User:	Symplectic Admin
Date Deposited:	27 Aug 2024 07:45
Last Modified:	22 May 2026 22:04
DOI:	10.1111/jns.12650
Open Access URL:	https://onlinelibrary.wiley.com/doi/10.1111/jns.12...
Related Websites:	Author Publisher
URI:	https://livrepository.liverpool.ac.uk/id/eprint/3184063
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