Burgess, J, Marshall, A, Rapteas, L, Hamill, KJ, Marshall, A, Malik, RA, Frank, B and Alam, U (2024) Automated immunohistochemistry of intra-epidermal nerve fibres in skin biopsies: A proof-of-concept study. Journal of the Peripheral Nervous System (JPNS), 29 (3). pp. 329-338. ISSN 1085-9489

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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.

Item Type:	Article
Uncontrolled Keywords:	Nerve Fibers; Epidermis; Skin; Humans; Ubiquitin Thiolesterase; Biopsy; Immunohistochemistry; Reproducibility of Results; Adult; Aged; Middle Aged; Female; Male; Small Fiber Neuropathy; Proof of Concept Study; automated; density; digital pathology; fibromyalgia syndrome; immunohistochemistry; intra‐epidermal nerve fibres; pathology; quantification; small fibre neuropathy; Humans; Nerve Fibers; Immunohistochemistry; Female; Male; Adult; Middle Aged; Proof of Concept Study; Skin; Small Fiber Neuropathy; Biopsy; Epidermis; Aged; Ubiquitin Thiolesterase; Reproducibility of Results; Neurodegenerative; Peripheral Neuropathy; Neurosciences; Humans; Nerve Fibers; Immunohistochemistry; Female; Male; Adult; Middle Aged; Proof of Concept Study; Skin; Small Fiber Neuropathy; Biopsy; Epidermis; Aged; Ubiquitin Thiolesterase; Reproducibility of Results; 1109 Neurosciences; Neurology & Neurosurgery
Subjects:	Q Science > QH Natural history > QH301 Biology Q Science > QH Natural history > QH426 Genetics
Divisions:	Psychology (from Sep 2019)
Publisher:	Wiley
SWORD Depositor:	A Symplectic
Date Deposited:	28 Nov 2024 18:23
Last Modified:	28 Nov 2024 18:30
DOI or ID number:	10.1111/jns.12650
URI:	https://researchonline.ljmu.ac.uk/id/eprint/24917

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