Challenges of automated screening and differentiation of non-organ specific autoantibodies on HEp-2 cells

Hiemann, Rico; Büttner, Thomas; Krieger, Thorsten; Roggenbuck, Dirk; Sack, Ulrich; Conrad, Karsten

doi:10.1016/j.autrev.2009.02.033

Cited by 146 publications

(134 citation statements)

References 19 publications

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“…The novel imaging-platform technology VideoScan (AKLIDES 1 ), which has recently been shown to be useful for the automated assessment of cell-based IIF assays in the serology of rheumatic diseases, was employed (13,21,22). There is a continuously growing need for cost efficient autoantibody testing technology that may combine both screening by IIF and multiplex evaluation of specific antibodies, on one platform.…”

Section: Discussionmentioning

confidence: 99%

“…There is a continuously growing need for cost efficient autoantibody testing technology that may combine both screening by IIF and multiplex evaluation of specific antibodies, on one platform. Current approaches for multiplex testing such as Luminex 1 or microarray technologies do not appear to be applicable in this context since they do not provide pattern recognition (13).…”

Section: Discussionmentioning

confidence: 99%

“…The assay characteristics and clinical value of the determination of ANA to six defined ENA are investigated and discussed, as an aid in the serological diagnosis of systemic rheumatic diseases. The interpretation system can also be used for the automated reading of HEp-2 cell-based IIF for the detection of ANA (13) and would be the first combined platform for both ANA screening and differentiation.…”

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confidence: 99%

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Multiplex assessment of non‐organ‐specific autoantibodies with a novel microbead‐based immunoassay

Großmann

Roggenbuck

Schröder³

et al. 2011

Cytometry Pt A

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Advances in immunofluorescence assay development paved the way for the simultaneous detection of several antibodies in one sample, for the serological diagnosis of systemic rheumatic diseases. Standardized automated screening of such antibodies can be achieved by HEp-2 cell-based indirect immunofluorescence (IIF) using a multicolor fluorescence imaging technical platform. To create a common platform for both screening and specific antibody assessment, multiplex measurement of antibodies using fluorescence-coded immobilized microbeads was employed on the same platform. The multicolor fluorescence detection system VideoScan (AKLIDES 1 ) was used for the fluorescence analysis of a multiplex microbead-based immunoassay (MIA). First, immunoglobulin G (IgG) was covalently coupled to one microbead population in duplicate and in three independent experiments. The coupled IgG was detected by a Cy TM 5-conjugated secondary antibody. Thus, intra-and interassay coefficients of variation (CV) were obtained. Second, a multiplex determination of antinuclear autoantibodies (ANA) to Scl-70, Sm, dsDNA, SS-A (Ro60), CENP-B, and La/SS-B by solid-phase MIA was investigated, using 72 sera from patients with autoimmune diseases such as systemic lupus erythematosus and systemic sclerosis (SS). The reproducibility study revealed intra-assay CVs ranging from 3.2% to 9.9%, and interassay CVs ranging from 9.6% to 14.7%. The detection of Scl-70-, Sm-, CENP-B-, and La/SS-B-ANA with MIA showed very good agreement with the ELISA results (kappa 5 1.0). The resulting relative sensitivities and specificities for Scl-70-, Sm-, CENP-B-, dsDNA-, and La/SS-B-ANA were 100%, respectively, with the exception of dsDNA (specificity 97%). Multiplex detection by immobilized fluorescence-coded microbeads using multicolor fluorescence is a reliable method for the assessment of rheumatic-disease-specific antibodies. Multicolor fluorescence analyses with pattern detection algorithms provide a common platform technique for both the screening of ANA by cell-based IIF and specific antibody assessment by multiplex detection. ' 2011 International Society for Advancement of Cytometry Key termsHEp-2 cell; multiplex; microbead-based immunoassay; autoantibodies ANTINUCLEAR antibodies (ANA) to extractable nuclear antigens (ENA) are hallmarks in the serological diagnosis of systemic rheumatic diseases (1). The detection of ANA by indirect immunofluorescence (IIF) employing HEp-2 cells as a multiple antigen source has been established as the ''gold standard'' in routine diagnostics (2). However, advances in assay development and recombinant technology have paved the way for the detection of ANA to individual ENA, such as double-stranded DNA (dsDNA), improving the diagnostic power of ANA testing. The growing variety of ANA found in different systemic rheumatic diseases has generated the need for innovative techniques to overcome the shortcomings of single ENA detection, with regard to cost and time taken to obtain results (3). Hence, multiplexed platforms have been ...

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Multiplex assessment of non‐organ‐specific autoantibodies with a novel microbead‐based immunoassay

Großmann

Roggenbuck

Schröder³

et al. 2011

Cytometry Pt A

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“…Some of the relevant biological applications of PREs at the cellular and subcellular levels include classification of cells and nuclei based on their morphological, textural, and appearance features (24,25), and interpreting and analyzing localization of proteins, antibodies, and subcellular structures within the cell (26)(27)(28)(29). For instance, Hill et al (30) assessed the impact of imperfect segmentation on the quality of highcontent screening data using a support vector machine (SVM)-based PRE to identify accurately delineated nuclei.…”

Section: Introductionmentioning

confidence: 99%

Automatic segmentation and supervised learning‐based selection of nuclei in cancer tissue images

et al. 2012

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Analysis of preferential localization of certain genes within the cell nuclei is emerging as a new technique for the diagnosis of breast cancer. Quantitation requires accurate segmentation of 100-200 cell nuclei in each tissue section to draw a statistically significant result. Thus, for large-scale analysis, manual processing is too time consuming and subjective. Fortuitously, acquired images generally contain many more nuclei than are needed for analysis. Therefore, we developed an integrated workflow that selects, following automatic segmentation, a subpopulation of accurately delineated nuclei for positioning of fluorescence in situ hybridization-labeled genes of interest. Segmentation was performed by a multistage watershed-based algorithm and screening by an artificial neural network-based pattern recognition engine. The performance of the workflow was quantified in terms of the fraction of automatically selected nuclei that were visually confirmed as well segmented and by the boundary accuracy of the well-segmented nuclei relative to a 2D dynamic programming-based reference segmentation method. Application of the method was demonstrated for discriminating normal and cancerous breast tissue sections based on the differential positioning of the HES5 gene. Automatic results agreed with manual analysis in 11 out of 14 cancers, all four normal cases, and all five noncancerous breast disease cases, thus showing the accuracy and robustness of the proposed approach. Published 2012 Wiley-Periodicals, Inc. y

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“…The assay is subjective as it relies heavily on human interpretation [8]. Heterogeneity in microscopes, light power, lens magnification and Hep-2, substantially contributes to its variability [9]. Its limitations can be minimized using automated microscopes, although not all ANA patterns can be recognized by such systems.…”

Section: Introductionmentioning

confidence: 99%

Predictive autoimmunity using autoantibodies: screening for anti-nuclear antibodies

Pérez

Gilburd

Cabrera-Marante³

et al. 2017

Clinical Chemistry and Laboratory Medicine (CCLM)

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Background: Early detection of antinuclear antibodies (ANA) in asymptomatic subjects is useful to predict autoimmune diseases years before diagnosis. ANA have been determined by indirect immunofluorescence (IIF) using human epithelial type 2 (HEp-2) cells, which is considered the gold standard technique. Multiplex technology (BioPlex ANA Screen) has been introduced for ANA evaluation in recent years. Nevertheless, concordance between BioPlex and IIF is low and there is no harmonization between both methods for detection of autoantibodies. This study has aimed to clarify the clinical significance of autoantibodies detected by BioPlex ANA Screen in subjects with undiagnosed clinical suspicion of autoimmune disease and to determine the predictive value of autoantibodies detected by BioPlex ANA Screen. Methods: A 3-year follow-up study was performed of 411 subjects without a clear diagnosis of autoimmune diseases in whom autoantibodies were detected by BioPlex ANA Screen that were negative by IIF on HEp-2 cells.

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Challenges of automated screening and differentiation of non-organ specific autoantibodies on HEp-2 cells

Cited by 146 publications

References 19 publications

Multiplex assessment of non‐organ‐specific autoantibodies with a novel microbead‐based immunoassay

Multiplex assessment of non‐organ‐specific autoantibodies with a novel microbead‐based immunoassay

Automatic segmentation and supervised learning‐based selection of nuclei in cancer tissue images

Predictive autoimmunity using autoantibodies: screening for anti-nuclear antibodies

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