Utility of immunofluorescence in dermatology

Shetty, Varsha M.; Subramaniam, Kumudhini; Rao, Raghavendra

doi:10.4103/2229-5178.198774

Cited by 24 publications

(3 citation statements)

References 31 publications

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“…The transport medium of choice is the immunofluorescence-specific solution, Michel's medium [43], which is widely available commercially and preserves immune complexes for up to 6 months at room temperature [44]. Following receipt in the laboratory, specimens are typically rinsed in phosphate buffered saline, to remove ammonium salts present in Michel's medium and then snap-frozen in an embedding compound such as OCT, following careful orientation of the specimen [45]. A series of frozen tissue sections, ideally of 4-5 μm thickness, are cut using a cryostat and placed on microscope slides, prior to addition of fluorescent conjugates and incubation, to facilitate immune complex formation.…”

Section: Direct Immunofluorescence (Dif)mentioning

confidence: 99%

Diagnostic Techniques in Autoimmune Blistering Diseases

Mee

2023

Br J Biomed Sci

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Autoimmune blistering diseases (AIBD) comprise a heterogeneous group of uncommon disorders of the skin and mucous membranes, characterised by antibodies targeting structural proteins within epithelial tissue and the underlying basement membrane. There can be significant overlap in clinical presentation of these diseases and accurate diagnosis relies on the detection and characterisation of relevant autoantibodies. Immunofluorescence provides the gold-standard diagnostic tool for these diseases, identifying both tissue-bound autoantibodies in biopsy material using direct immunofluorescence and circulating antibodies in serum through indirect immunofluorescence. Following advances in the identification and subsequent characterisation of numerous antigenic targets in these diseases, the development of antigen-specific tests, in particular, enzyme-linked immunosorbent assays on serum specimens, has provided a third key tool to not only identify, but also quantify AIBD autoantibodies. This quantification has proven particularly useful in monitoring disease activity and informing clinical management decisions. Accurate diagnosis of these diseases is important since optimal treatment strategies differ between them and, prognostically, some diagnoses are associated with an increased risk of malignancy. This review outlines the molecular pathology underlying the major AIBD and describes how the three principal techniques can be used in combination, to provide best practice for diagnosis and treatment monitoring.

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Section: Direct Immunofluorescence (Dif)mentioning

confidence: 99%

Diagnostic Techniques in Autoimmune Blistering Diseases

Mee

2023

Br J Biomed Sci

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“…In practice, light microscopic findings, combined with direct immunofluorescence (DIF) results, are used for a definitive pathological diagnosis [4,5].…”

Section: Introductionmentioning

confidence: 99%

“…DIF microscopy is considered the gold standard for differentiating various vesiculobullous diseases [6,7]. In addition to diagnosis, DIF also aids in monitoring response to therapy and predicting relapse [3,5].…”

Section: Introductionmentioning

confidence: 99%

Utility of Direct Immunofluorescence in Cutaneous Autoimmune Bullous Disorders

Brar¹,

Sharma²,

Nauhria³

et al. 2021

Cureus

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BackgroundAutoimmune bullous disorders (AIBD) are a heterogeneous group of disorders with substantial clinical overlap associated with blistering of skin or mucosa. AimsThe present study aimed to study the histopathological spectrum and evaluate the utility of direct immunofluorescence (DIF) on snap-frozen and paraffin-embedded sections in resolving the differential diagnosis of AIBD and connective tissue disorders of the skin. We also compared the efficacy of DIF on paraffin versus the snap-frozen sections in diagnosing AIBD. MethodsThe present study was conducted for three years (2017-2019) and included 27 biopsies. We also included a retrospective analysis that included 25 biopsies collected over three years (2014)(2015)(2016)(2017). Histopathological examination and DIF were conducted on all samples. ResultsPemphigus vulgaris was the most common autoimmune cutaneous disorder constituting 37% (n = 10) in prospective and 36% (n = 9) in the retrospective study. DIF showed a specificity of 81.25% in our prospective study. While on the paraffin-embedded sections, it showed a specificity of 66.6% in our retrospective study. In the prospective study, DIF on paraffin-embedded sections had a positivity rate of 43.75% as compared to 81.25% in DIF done on snap-frozen sections. ConclusionDIF is a sensitive tool for the diagnosis as well as distinguishing immune-mediated bullous disorders from other lesions primarily when performed on snap-frozen sections. The diagnostic yield is enhanced by DIF in cases that pose a diagnostic dilemma both clinically and histologically. The final diagnosis depends on all clinical, histopathological and immunofluorescence findings.

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Histopathology of the Skin: General Principles

Calonje,

Mee

2024

Rook's Textbook of Dermatology

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Utility of immunofluorescence in dermatology

Cited by 24 publications

References 31 publications

Diagnostic Techniques in Autoimmune Blistering Diseases

Diagnostic Techniques in Autoimmune Blistering Diseases

Utility of Direct Immunofluorescence in Cutaneous Autoimmune Bullous Disorders

Histopathology of the Skin: General Principles

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