Automated direct immunofluorescence analyses of skin biopsies

Lemcke, Susanne; Sokolowski, Siegmar; Rieckhoff, Nicole; Buschtez, Michael; Kaffka, Christian; Winter-Keil, Antje; Schaller, Christine; Rottmann, Norbert; Sadik, Christian D.; Stöcker, W.; Zillikens, Detlef; Schmidt, Enno

doi:10.1111/cup.12637

Cited by 9 publications

(8 citation statements)

References 17 publications

(19 reference statements)

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“…Second, if substrates for IF microscopy or ELISA are not available, biochips spotted with primate salt-split skin, recombinant BP180 and BP230 expressing cells can be purchased and used for indirect IF microscopy for the detection of circulating autoantibodies directed against BP180 or BP230 ( 101 ). This may be even automated, for high-throughput analysis ( 102 , 103 ).…”

Section: Putative Explanations For This Surgementioning

confidence: 99%

The Growing Incidence of Bullous Pemphigoid: Overview and Potential Explanations

2018

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Bullous pemphigoid (BP) is the most common type of subepidermal autoimmune bullous diseases. BP characteristically affects the elderly and is seen mainly in patients older than 70 years. While the annual incidence of BP has been estimated to be between 2.4 and 23 cases per million in the general population, it rises exponentially to 190–312 cases per million in individuals older than 80 years. In addition, a growing body of evidence reports a remarkable trend of increased incidence of BP, showing a 1.9- to 4.3-fold rise over the past two decades. This demonstrable increase warrants a higher awareness of the increased risk to develop BP. This review summarizes the current understanding of the epidemiological features of BP and sheds light on the putative explanations for its growing incidence.

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Section: Putative Explanations For This Surgementioning

confidence: 99%

The Growing Incidence of Bullous Pemphigoid: Overview and Potential Explanations

2018

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“…In dermatitis herpetiformis, DIF microscopy reveals granular deposition of IgA at the dermal papillae and along the DEJ. An automated staining for DIF microscopy sections has recently been developed and revealed more intensive IF staining and reduced background compared to the manual procedure due to continuous movement and overhead incubation ( 40 ). A flowchart navigating through differential diagnoses using DIF microscopy is shown in Figure 1 .…”

Section: Direct Immunofluorescence Microscopymentioning

confidence: 99%

Diagnosis of Autoimmune Blistering Diseases

2018

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Autoimmune skin blistering diseases (AIBD) are characterized by autoantibodies that are directed against structural proteins in the skin and adjacent mucous membranes. Some clinical signs are typical for a specific AIBD, however, correct diagnosis requires the detection of tissue-bound or circulating autoantibodies. The gold standard for diagnosis of AIBD is the detection of autoantibodies or complement component 3 by direct immunofluorescence (DIF) microscopy of a perilesional biopsy. Circulating antibodies can be detected via indirect immunofluorescence (IIF) microscopy of different tissue substrates including human skin, monkey esophagus, and more recently, recombinant forms of the different target antigens. Latter are also employed in various commercial ELISA systems and by immunoblotting in in-house assays available in specialized laboratories. ELISA systems are also particularly valuable for monitoring of the disease activity during the disease course which can be helpful for treatment decisions. Exact diagnosis is essential for both treatment and prognosis, since some AIBD are associated with malign tumors such as paraneoplastic pemphigus and anti-laminin 332 mucous membrane pemphigoid. This review presents clinical and immunopathological features of AIBD for the state-of the art diagnosis of these disorders.

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“…Nevertheless, more recently a new technology for automation and standardization of DIF staining was developed. Lemcke et al () indicated that some DIF disadvantages (inconsistent staining results and unspecific background) independent on the chosen techniques may be solved with presented automated system. Thus, perhaps an automated system of DIF improves its diagnostic accuracy.…”

Section: Discussionmentioning

confidence: 99%

Blue light‐emitting diode technology‐operated microscopy is preferable to both short arc mercury lamp‐operated microscopy and laser scanning confocal microscopy for direct immunofluorescence images evaluation in routinely diagnosing subepidermal autoimmune blistering diseases

Gornowicz‐Porowska

Bowszyc‐Dmochowska

Raptis-Bolwach

et al. 2019

Microscopy Res & Technique

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Direct immunofluorescence (DIF) microscopy still is the gold standard in diagnosing and differentiating subepidermal autoimmune blistering diseases (SABDs) from other bullous diseases. New optical systems were developed that aim to facilitate DIF images evaluation. The aim of the study was to evaluate the usefulness of three fluorescence microscopy systems with different light source for routine diagnostics of SABDs with DIF. In total, perilesional tissue samples for DIF from 34 SABD patients were examined under the three commercial microscopy systems (short arc mercury lamp‐operated microscopy—MLM, blue light‐emitting diode technology‐operated microscopy—bLED and laser scanning confocal microscopy—LSCM) for the detection and pattern analysis of IgA, IgG, IgG1, IgG4, C3 immunoreactants along the dermal–epidermal junction (DEJ) by three independent observers simultaneously. MLM, bLED, and LSCM provided comparable quality of disease‐characterizing immunoreactants imaging (number of immunoreactant types detected and patterns of their deposition along DEJ were the same) but screening of slides was quicker using bLED than both LSCM and MLM, as statistical analysis indicated. It is concluded that bLED is an efficient and preferable system for routinely diagnosing SABDs cost‐effectively. Research Highlights New optical systems were developed that aim to facilitate direct immunofluorescence evaluation. Here, we evaluate the usefulness of three fluorescence microscopy systems with different light source for routine diagnostics concluding that that bLED is an efficient and preferable system.

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Automated direct immunofluorescence analyses of skin biopsies

Abstract: EUROTide/EUROPath applied in an automated system improves diagnostic accuracy and saves reagents. Larger studies in other routine laboratories may further explore the value of the EUROTide/EUROPath technology and may include comparison with other automated stainers.

Cited by 9 publications

References 17 publications

The Growing Incidence of Bullous Pemphigoid: Overview and Potential Explanations

The Growing Incidence of Bullous Pemphigoid: Overview and Potential Explanations

Diagnosis of Autoimmune Blistering Diseases

Blue light‐emitting diode technology‐operated microscopy is preferable to both short arc mercury lamp‐operated microscopy and laser scanning confocal microscopy for direct immunofluorescence images evaluation in routinely diagnosing subepidermal autoimmune blistering diseases

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