Der CytoBead-Assay – Eine neue Möglichkeit der multiparametrischen Autoantikörperanalytik bei systemischen Autoimmunerkrankungen

Sowa, Mandy; Großmann, Knut; Scholz, Juliane; Röber, Nadja; Rödiger, Stefan; Schierack, Peter; Conrad, Karsten; Roggenbuck, Dirk; Hiemann, Rico

doi:10.1515/labmed-2014-0041

Cited by 2 publications

(3 citation statements)

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“…Thus, there is obviously a demand for multiplex methods to analyze CD-specific autoAb and IgA deficiency simultaneously in one sample. To the best of our knowledge, the CytoBead technique based on digital fluorescence is the first assay technology available so far which provides the unique opportunity to combine autoAb testing employing tissue and purified antigenic targets in one reaction environment [ 28 ]. Thus, we developed the CytoBead CeliAK assay for the simultaneous detection of CD-specific IgA autoAb including the reference autoAb EmA and total IgA testing using this novel technology.…”

Section: Discussionmentioning

confidence: 99%

“…Thus, we developed the CytoBead CeliAK assay for the simultaneous detection of CD-specific IgA autoAb including the reference autoAb EmA and total IgA testing using this novel technology. The efficacy thereof has been demonstrated for similar laboratory demands of multiplex autoAb testing like antinuclear and antineutrophilic cytoplasmic Ab analyses recently [ 27 , 28 , 32 ]. Novel pattern recognition algorithms and digital fluorescence have paved the way for quantification and automation of autoAb testing through IIF interpretation systems [ 33 – 36 ].…”

Section: Discussionmentioning

confidence: 99%

“…Consequently, taking into account the number of tests required for an appropriate CD serology, there is a need for new strategies to reduce the workload and laboratory costs in routine laboratories [ 23 – 26 ]. Thus, the aim of this study was to evaluate a multiplex strategy detecting CD-specific IgA autoAbs with the novel CytoBead technique which has been already employed successfully for multiplex autoAb analysis [ 27 , 28 ]. The assay allows the simultaneous analysis of EmA, TG2autoAb, and DGAb as well as the determination of total IgA in one reaction environment by IIF for the first time.…”

Section: Introductionmentioning

confidence: 99%

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Simultaneous detection of celiac disease-specific IgA antibodies and total IgA

Großmann

Röber

Hiemann

et al. 2016

Autoimmun Highlights

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PurposeCeliac disease (CD) serology requires analysis of tissue transglutaminase type-2 (TG2autoAbs), deamidated gliadin (DGAbs), and as reference endomysial autoantibodies (EmA). Total IgA assessment helps to determine IgA-deficient CD patients. The novel multiplex indirect immunofluorescence (IIF) technique CytoBead was used to develop the first quantitative one-step serological CD assay comprising both simultaneous IgA autoAb and total IgA testing.MethodsCytoBead CeliAK detecting TG2autoAb, DGAb, EmA, and simultaneously total IgA uses fluorescent microparticles for antigen and antibody immobilization along with monkey-esophagus tissue sections on glass slides. The assay was interpreted visually by classical fluorescent microscopy and digital IIF using AKLIDES®. Overall, 380 samples (155 CD patients, 5 with IgA deficiency, 68 with cystic fibrosis, 59 with eye disease, 93 blood donors) were run for performance analysis. Data were compared with classical IgA autoAb analysis by ELISA and IIF.ResultsComparing CD-specific IgA autoAb testing by CytoBead with classical IIF and ELISA, very good agreements for EmA, TG2autoAb, and DGAb were determined (Cohen’s κ = 0.98, 0.96, 0.85, respectively). The difference between multiplex and single testing revealed a significant difference for TG2autoAb testing only (McNemar, p = 0.0078). Four CD patients and 4 controls demonstrated TG2autoAb positivity by ELISA but were negative by CytoBead. Further, 140/155 (90.9 %) CD patients demonstrated TG2autoAb levels above ten times the upper normal and all five IgA-deficient samples IgA levels <0.2 g/L by CytoBead.ConclusionsThe novel multiplex CytoBead CeliAK enables simultaneous CD-specific autoAb and IgA deficiency analyses comparable with classical testing by single-parameter assays. Thus, comprehensive CD serology by CytoBead can alleviate the workload in routine laboratories.Electronic supplementary materialThe online version of this article (doi:10.1007/s13317-016-0073-2) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Simultaneous detection of celiac disease-specific IgA antibodies and total IgA

Großmann

Röber

Hiemann

et al. 2016

Autoimmun Highlights

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Close Spectroscopic Look at Dye-Stained Polymer Microbeads

et al. 2018

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Dye-stained micrometer-sized polymer beads are important tools in the life sciences with applications in biomedical, biochemical, and clinical research. Here, bead-based assays are increasingly used, for example, in DNA sequencing and the detection of autoimmune diseases or pathogenic microorganisms. Moreover, stained beads are employed as calibration tools for fluorescence microscopy and flow cytometry methods with increasing complexity. To address the requirements concerning the relevant fluorescence features, the spectroscopic properties of representative polymer beads with diameters ranging from about 1 to 10 μm stained with varying concentrations of rhodamine 6G were systematically assessed. The observed dependence of the spectral properties, fluorescence decay kinetics, and fluorescence quantum yields on bead size and dye loading concentration is attributed to different fluorescence characteristics of fluorophores located in the particle core and near-surface dye molecules. Supported by the fluorescence anisotropy measurements, the origin of the observed alteration of fluorescence features is ascribed to a combination of excitation energy transfer and polarity-related effects that are especially pronounced at the interface of the bead and the surrounding medium. The results of our studies underline the need to carefully control and optimize all parameters that can affect the fluorescence properties of the dye-stained beads.

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Der CytoBead-Assay – Eine neue Möglichkeit der multiparametrischen Autoantikörperanalytik bei systemischen Autoimmunerkrankungen

Cited by 2 publications

References 41 publications

Simultaneous detection of celiac disease-specific IgA antibodies and total IgA

Simultaneous detection of celiac disease-specific IgA antibodies and total IgA

Close Spectroscopic Look at Dye-Stained Polymer Microbeads

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