TY - GEN A1 - Hiemann, Rico A1 - Hilger, Nadja A1 - Michel, Jörg A1 - Nitschke, Jörg A1 - Böhm, Alexander A1 - Anderer, Ursula A1 - Weigert, Martin A1 - Sack, Ulrich T1 - Automatic Analysis of Immunofluorescence Patterns of HEp‐2 Cells T2 - Annals of the New York Academy of Sciences N2 - The standard screening test for the recognition of autoimmune diseases is the proof of autoantibodies in serum of patients by indirect immunofluorescence (IIF) based on HEp‐2 cells. Manual evaluation of this test is very subjective, slow, and there are no objective parameters as guidelines available. Interlaboratory tests showed occasionally large deviations in the test evaluation resulting in a high variance of results. The aim of this project is fast, objective, safe, and economical automatic analysis of HEp‐2 IIF patterns. Images of IIF patterns were completely and automatically captured using an inverse motorized fluorescence microscope. Thereby, device‐specific parameters were controlled automatically, too. For fast analysis of IIF patterns new algorithms of image processing were developed. Artifacts were recognized and excluded from analysis by the developed software. Analysis of more than 80,000 images clearly demonstrated full automatization and fast processing of IIF patterns. Additionally serum‐specific fluorescence could be easily distinguished from background. Even very weak but positive patterns can be recognized and used for diagnosis. A detailed separation into different basic patterns is possible. Objective, fast, and disease‐related economical analysis of HEp‐2 immunofluorescence patterns is feasible. The implemented software algorithms allowed a mathematical way of describing IIF patterns and can therefore be a useful tool for the needed standardization process. KW - Autoimmune diagnostics KW - HEp-2 cell KW - Immunofluorescence KW - Automatic analysis KW - Screening Y1 - 2007 U6 - https://doi.org/10.1196/annals.1398.042 SN - 0077-8923 VL - 1109 IS - 1 SP - 358 EP - 371 ER - TY - GEN A1 - Frömmel, Ulrike A1 - Lehmann, Werner A1 - Rödiger, Stefan A1 - Böhm, Alexander A1 - Nitschke, Jörg A1 - Weinreich, Jörg A1 - Groß, Julia A1 - Roggenbuck, Dirk A1 - Zinke, Olaf A1 - Ansorge, Hermann A1 - Vogel, Steffen A1 - Klemm, Per A1 - Wex, Thomas A1 - Schröder, Christian A1 - Wieler, Lothar H. A1 - Schierack, Peter T1 - Adhesion of human and animal Escherichia coli strains in association with their virulence-associated genes and phylogenetic origins T2 - Applied and environmental microbiology N2 - Intestinal colonization is influenced by the ability of the bacterium to inhabit a niche, which is based on the expression of colonization factors. Escherichia coli carries a broad range of virulence-associated genes (VAGs) which contribute to intestinal (inVAGs) and extraintestinal (exVAGs) infection. Moreover, initial evidence indicates that inVAGs and exVAGs support intestinal colonization. We developed new screening tools to genotypically and phenotypically characterize E. coli isolates originating in humans, domestic pigs, and 17 wild mammal and avian species. We analyzed 317 isolates for the occurrence of 44 VAGs using a novel multiplex PCR microbead assay (MPMA) and for adhesion to four epithelial cell lines using a new adhesion assay. We correlated data for the definition of new adhesion genes. inVAGs were identified only sporadically, particularly in roe deer (Capreolus capreolus) and the European hedgehog ( Erinaceus europaeus). The prevalence of exVAGs depended on isolation from a specific host. Human uropathogenic E. coli isolates carried exVAGs with the highest prevalence, followed by badger (Meles meles) and roe deer isolates. Adhesion was found to be very diverse. Adhesion was specific to cells, host, and tissue, though it was also unspecific. Occurrence of the following VAGs was associated with a higher rate of adhesion to one or more cell lines: afa-dra, daaD, tsh, vat, ibeA, fyuA, mat, sfa-foc, malX, pic, irp2, and papC. In summary, we established new screening methods which enabled us to characterize large numbers of E. coli isolates. We defined reservoirs for potential pathogenic E. coli. We also identified a very broad range of colonization strategies and defined potential new adhesion genes. Y1 - 2013 U6 - https://doi.org/10.1128/AEM.01384-13 SN - 1098-5336 SN - 0099-2240 VL - 79 IS - 19 SP - 5814 EP - 5829 ER - TY - GEN A1 - Liebsch, Claudia A1 - Rödiger, Stefan A1 - Böhm, Alexander A1 - Nitschke, Jörg A1 - Weinreich, Jörg A1 - Fruth, Angelika A1 - Roggenbuck, Dirk A1 - Lehmann, Werner A1 - Schedler, Uwe A1 - Juretzek, Thomas A1 - Schierack, Peter T1 - Solid-phase microbead array for multiplex O-serotyping of Escherichia coli T2 - Microchimica Acta Y1 - 2017 U6 - https://doi.org/10.1007/s00604-017-2088-4 SN - 0026-3672 SN - 1436-5073 VL - 184 IS - 5 SP - 1405 EP - 1415 ER - TY - GEN A1 - Kolenda, Rafał A1 - Burdukiewicz, Michał A1 - Schiebel, Juliane A1 - Rödiger, Stefan A1 - Sauer, Lysann A1 - Szabo, Istvan A1 - Orlowska, Aleksandra A1 - Weinreich, Jörg A1 - Nitschke, Jörg A1 - Böhm, Alexander A1 - Gerber, Ulrike A1 - Roggenbuck, Dirk A1 - Schierack, Peter T1 - Adhesion of Salmonella to Pancreatic Secretory Granule Membrane Major Glycoprotein GP2 of Human and Porcine Origin Depends on FimH Sequence Variation T2 - Frontiers in Microbiology Y1 - 2018 U6 - https://doi.org/10.3389/fmicb.2018.01905 SN - 1664-302X VL - 9 IS - 1905 ER - TY - GEN A1 - Schiebel, Juliane A1 - Böhm, Alexander A1 - Nitschke, Jörg A1 - Burdukiewicz, Michał A1 - Weinreich, Jörg A1 - Ali, Aamir A1 - Roggenbuck, Dirk A1 - Rödiger, Stefan A1 - Schierack, Peter T1 - Genotypic and phenotypic characteristics in association with biofilm formation in different pathotypes of human clinical Escherichia coli isolates T2 - Applied and Environmental Microbiology Y1 - 2017 U6 - https://doi.org/10.1128/AEM.01660-17 SN - 1098-5336 SN - 0099-2240 VL - 83 IS - 24 ER - TY - GEN A1 - Frömmel, Ulrike A1 - Böhm, Alexander A1 - Nitschke, Jörg A1 - Weinreich, Jörg A1 - Groß, Julia A1 - Rödiger, Stefan A1 - Wex, Thomas A1 - Ansorge, Hermann A1 - Zinke, Olaf A1 - Schröder, Christian A1 - Roggenbuck, Dirk A1 - Schierack, Peter T1 - Adhesion patterns of commensal and pathogenic Escherichia coli from humans and wild animals on human and porcine epithelial cell lines T2 - Gut Pathogens N2 - Abstract BACKGROUND: Different strategies of colonization or infection by E. coli result in formation of certain adhesion patterns which help also in classifying intestinal E. coli into pathotypes. Little is known about adhesion patterns and host- and tissue adaption of commensal E. coli and about E. coli originating in clinically healthy hosts carrying pathotype-specific virulence-associated genes. FINDINGS: Adhesion pattern of E. coli (n = 282) from humans and from 18 animal species were verified on intestinal human Caco-2 and porcine IPEC-J2 cells and, furthermore, for comparison on human urinary bladder 5637, porcine kidney PK-15 epithelial and HEp-2 cells. The analysis was carried out on 150,000 images of adhesion assays.Adhesion patterns were very diverse; 88 isolates were completely non-adherent, whereas 194 adhered to at least one cell line with the dominant adhesion patterns "diffusely distributed" and "microcolony formation". Adhesion patterns "chains" and "clumps" were also visible. Chain formation was mediated by the presence of epithelial cells. Clump formation was very specific on only the 5637 cell line. All enteropathogenic (eae+) E. coli (EPEC; n = 14) were able to form microcolonies which was cell line specific for each isolate. Most EPEC formed microcolonies on intestinal IPEC-J2 and Caco-2 but several also on urinary tract cells. Shigatoxin-producing (stx+) E. coli (n = 10) showed no specific adhesion patterns. CONCLUSIONS: E. coli isolates were highly diverse. Commensal and pathogenic isolates can adhere in various forms, including diffuse distribution, microcolonies, chains and clumps. Microcolony formation seems to be a global adhesion strategy also for commensal E. coli. Y1 - 2013 U6 - https://doi.org/10.1186/1757-4749-5-31 SN - 1757-4749 VL - 5 IS - 31 ER - TY - CHAP A1 - Kolenda, Rafał A1 - Burdukiewicz, Michał A1 - Szabo, M. A1 - Orlowska, Aleksandra A1 - Nitschke, Jörg A1 - Böhm, Alexander A1 - Schierack, Peter T1 - Influence of Salmonella FimH sequence variation on binding to pancreatic secretory granule membrane major glycoprotein GP2 of human and porcine origin T2 - 5th ASM Conference on Salmonella, Potsdam, 29.08-01.09.2016 Y1 - 2016 UR - https://www.asm.org/images/1903%20asm%20salmonella%20program%20web3.pdf SP - S. 125 PB - American Society for Microbiology CY - Washington, DC ER - TY - GEN A1 - Awan, Asad Bashir A1 - Schiebel, Juliane A1 - Böhm, Alexander A1 - Nitschke, Jörg A1 - Sarwar, Yasra A1 - Schierack, Peter A1 - Ali, Aamir T1 - Association of biofilm formation and cytotoxic potential with multidrug resistance in clinical isolates of Pseudomonas aeruginosa T2 - EXCLI journal Y1 - 2019 SN - 1611-2156 VL - 18 SP - 79 EP - 90 ER - TY - GEN A1 - Rödiger, Stefan A1 - Lehmann, Werner A1 - Frömmel, Ulrike A1 - Böhm, Alexander A1 - Nitschke, Jörg A1 - Burdukiewicz, Michał A1 - Schröder, Christian A1 - Dangla, Rémi A1 - Droniou, Magali A1 - Schierack, Peter T1 - The Modi Operandi of the VideoScan Platform for the Detection and Analysis of Nucleic Acids. Life Science Day 2013 & 17. Leibniz Conference of Advanced Science, Freie Universität Berlin, 24.10.2013 Y1 - 2013 UR - https://leibniz-institut.de/Konferenzen/lsd2013/roediger_the_modi_operandi_of_the_videoscan_platform.pdf ER - TY - CHAP A1 - Rödiger, Stefan A1 - Böhm, Alexander A1 - Nitschke, Jörg A1 - Stavitskaya, Luba A1 - Gruner, Melanie A1 - Kundzer, Alena V. A1 - Volkova, Margarita V. A1 - Generalov, I. A1 - Schmidt, Carsten A1 - Schröder, Christian A1 - Roggenbuck, Dirk A1 - Schierack, Peter T1 - Development of a Method for Multiplex Real-Time Analysis of Enzymatic Activity on a Microbead-Chip T2 - Infection, tumors and autoimmunity, report on the 11th Dresden Symposium on Autoantibodies held in Dresden on September 1-4, 2013 Y1 - 2013 SN - 978-3-89967-881-9 SP - 275 EP - 280 PB - Pabst Science Publ. CY - Lengerich [u.a.] ER - TY - CHAP A1 - Kolenda, Rafał A1 - Burdukiewicz, Michał A1 - Schiebel, Juliane A1 - Szabo, Istvan A1 - Orlowska, Aleksandra A1 - Nitschke, Jörg A1 - Böhm, Alexander A1 - Schierack, Peter T1 - Adhesion of Salmonella to pancreatic secretory granule membrane major glycoprotein GP2 depends on FimH sequence variation T2 - Abstractbook, Microbiology and Infection, 5th Joint Conference of the DGHM & VAAM, VAAM Annual Meeting 2017, 69th Annual Meeting of the DGHM Y1 - 2017 PB - Spektrumverlag CY - Heidelberg ER - TY - CHAP A1 - Burdukiewicz, Michał A1 - Mikolajczyk, A. A1 - Szabo, Istvan A1 - Nitschke, Jörg A1 - Böhm, Alexander A1 - Schierack, Peter A1 - Kolenda, Rafał T1 - Automated enumeration of Salmonella spp. in cell line infection assays and screening of cell line infectivity among various Salmonella isolates T2 - 5th ASM Conference on Salmonella, Potsdam, 29.08-01.09.2016 Y1 - 2016 UR - https://www.asm.org/images/1903%20asm%20salmonella%20program%20web3.pdf SP - S. 127 PB - American Society for Microbiology CY - Washington, DC ER - TY - CHAP A1 - Rödiger, Stefan A1 - Schierack, Peter A1 - Böhm, Alexander A1 - Nitschke, Jörg A1 - Berger, Ingo A1 - Frömmel, Ulrike A1 - Schmidt, Carsten A1 - Ruhland, Mirko A1 - Schimke, Ingolf A1 - Roggenbuck, Dirk A1 - Lehmann, Werner A1 - Schröder, Christian ED - Seitz, Harald ED - Berger, Ingo T1 - A highly versatile microscope imaging technology platform for the multiplex real-time detection of biomolecules and autoimmune antibodies T2 - Molecular diagnostics Y1 - 2013 SN - 978-3-642-37690-0 SN - 978-3-642-37691-7 SP - 35 EP - 74 PB - Springer CY - Berlin [u.a.] ER - TY - GEN A1 - Schiebel, Juliane A1 - Noack, Jonas A1 - Rödiger, Stefan A1 - Kammel, Anne A1 - Menzel, Friederike A1 - Schwibbert, Karin A1 - Weise, Matthias A1 - Weiss, Romano A1 - Böhm, Alexander A1 - Nitschke, Jörg A1 - Elimport, Alexey A1 - Roggenbuck, Dirk A1 - Schierack, Peter T1 - Analysis of three-dimensional biofilms on different material surfaces T2 - Biomaterials Science Y1 - 2020 U6 - https://doi.org/10.1039/D0BM00455C SN - 2047-4849 VL - 8 IS - 12 SP - 3500 EP - 3510 ER - TY - GEN A1 - Rödiger, Stefan A1 - Ruhland, Mirko A1 - Schröder, Christian A1 - Großmann, Kai A1 - Böhm, Alexander A1 - Nitschke, Jörg A1 - Berger, Ingo A1 - Schimke, Ingolf A1 - Schierack, Peter T1 - Fluorescence dye adsorption assay to quantify carboxyl groups on the surface of poly(methyl methacrylate) microbeads T2 - Analytical Chemistry Y1 - 2011 U6 - https://doi.org/10.1021/ac103277s SN - 0003-2700 SN - 1520-6882 VL - 83 IS - 9 SP - 3379 EP - 3385 ER - TY - CHAP A1 - Rödiger, Stefan A1 - Böhm, Alexander A1 - Nitschke, Jörg A1 - Stavitskaya, Luba A1 - Gruner, Melanie A1 - Kundzer, Alena V. A1 - Volkova, Margarita V. A1 - Generalov, I. A1 - Schmidt, Carsten A1 - Schröder, Christian A1 - Roggenbuck, Dirk A1 - Schierack, Peter T1 - Development of a Method for Multiplex Real-Time Analysis of Enzymati c Activity on a Microbead-Chip. T2 - Infections, Tumors and Autoimmunity, Report on the 11th Dresden Symposium on Autoantibodies held in Dresden on September 01. - 04., 2013 Y1 - 2013 SN - 978-3-89967-881-9 SP - 280 PB - Pabst Science Publ. CY - Lengerich ER -