Quality assurance for genotyping and resistance testing of Clostridium (Clostridioides) difficile isolates - Experiences from the first inter-laboratory ring trial in four German speaking countries

Affiliation
German National Reference Center for Clostridioides (Clostridium) difficile, Germany
Berger, Fabian K.;
Affiliation
German National Reference Center for Clostridioides (Clostridium) difficile, Germany
Mellmann, Alexander;
Affiliation
German National Reference Center for Clostridioides (Clostridium) difficile, Germany
von Müller, Lutz;
Affiliation
German National Reference Center for Clostridioides (Clostridium) difficile, Germany
Bischoff, Markus;
Affiliation
German National Reference Center for Clostridioides (Clostridium) difficile, Germany
Gärtner, Barbara C.; German speaking C. difficile laboratory study group:;
Affiliation
Thuringian State Authority for Consumer Protection (TLV), Bad Langensalza, Germany
Schneeberg, Alexander;
Affiliation
Clinical Microbiology, University Hospital Basel, Basel, Switzerland
Egli, Adrian;
Affiliation
German Federal Institute for Risk Assessment, Department Biological Safety, Berlin, Germany
Maurischat, Sven;
Affiliation
University of Cologne, Institute for Medical Microbiology, Immunology and Hygiene, Cologne, Germany
Jazmati, Nathalie;
Affiliation
Institut für Hygiene, Kliniken der Stadt Köln, Cologne, Germany
Malecki, Monika;
Affiliation
Leibniz-Institut DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen), Braunschweig, Germany
Nübel, Ulrich;
Affiliation
Leibniz-Institut DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen), Braunschweig, Germany
Junker, Vera;
Affiliation
Zentrum für Gesundheitsschutz-, Fachgebiet: Hygiene in Gesundheits- und Gemeinschaftseinrichtungen, Infektiologische Diagnostik, Dillenburg, Germany
Bettge-Weller, Gudrun;
Affiliation
MVZ Dr. Eberhard & Partner Dortmund (ÜBAG), Bereich Mikrobiologie, Dortmund, Germany
Pranada, Felix;
Affiliation
MVZ Dr. Eberhard & Partner Dortmund (ÜBAG), Bereich Mikrobiologie, Dortmund, Germany
Pranada, Arthur;
Affiliation
Laboratoire National de Santé, Service MycoBac-ARH, Dudelange, Luxembourg
Perrin, Monique;
Affiliation
Institut für Medizinische Mikrobiologie und Krankenhaushygiene, Universitätsklinikum Düsseldorf, Heinrich-Heine Universität, Düsseldorf, Germany
Kolbe-Busch, Susanne;
Affiliation
Institut für Medizinische Mikrobiologie und Krankenhaushygiene, Universitätsklinikum Düsseldorf, Heinrich-Heine Universität, Düsseldorf, Germany
MacKenzie, Colin;
Affiliation
Institut für Medizinische Mikrobiologie, Universitätsmedizin Göttingen, Göttingen, Germany
Groß, Uwe;
Affiliation
Institute of Medical Microbiology, Infection Control and Prevention, Otto-von-Guericke University of Magdeburg, Magdeburg, Germany
Färber, Jacqueline;
Affiliation
1 Abbott (Alere Technologies GmbH), Jena, Germany
Gawlik, Darius;
Affiliation
Institut für Medizinische Mikrobiologie und Hygiene, Dresden, Germany
Monecke, Stefan;
GND
12238136X
Seyboldt, Christian;
Affiliation
Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit (LGL) (Bavarian Health and Food Safety Authority), Sachgebiet GE2.2 Humanbakteriologie und Mykologie, Oberschleißheim, Germany
Berger, Anja;
Affiliation
Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit (LGL) (Bavarian Health and Food Safety Authority), Sachgebiet GE 1: Hygiene, Oberschleißheim, Germany
Tuschak, Christian;
Affiliation
Institute for Hygiene and Microbiology, University of Wuerzburg, Wuerzburg, Germany
Lâm, Thien-Tri;
Affiliation
Institut für Klinische Mikrobiologie und Hygiene Universitätsklinikum Regensburg, Regensburg, Germany
Hiergeist, Andreas;
Affiliation
Institut für Klinische Mikrobiologie und Hygiene Universitätsklinikum Regensburg, Regensburg, Germany
Reischl, Udo;
Affiliation
Landesgesundheitsamt Baden-Württemberg Ref. 93: Allgemeine Hygiene & Infektionsschutz, Stuttgart, Germany
Adam, Maja;
Affiliation
Institut für medizinische Mikrobiologie und Hygiene - Zentrum für anthropogene Infektionen Abteilung Klinische Molekularbiologie, Geschäftsfeld Öffentliche Gesundheit AGES - Österreichische Agentur für Gesundheit und Ernährungssicherheit GmbH, Austria
Baumgartner, Stefanie;
Affiliation
Institut für medizinische Mikrobiologie und Hygiene - Zentrum für anthropogene Infektionen Abteilung Klinische Molekularbiologie, Geschäftsfeld Öffentliche Gesundheit AGES - Österreichische Agentur für Gesundheit und Ernährungssicherheit GmbH, Austria
Mantlik, Katrin

Clostridium (Clostridioides) difficile is a major cause of nosocomial diarrhoea. A first inter-laboratory ring trial was performed in four European countries to evaluate the genotyping and antibiotic susceptibility testing (AST) accuracy. Six C. difficile isolates representing the epidemiologic important ribotypes (RT), RT001, RT002, RT010, RT014, RT027, and RT078 were blinded and send to 21 participating laboratories. Participants tested the samples with their genotyping and AST methods in use for concordance with reference. A total of 21 genotyping- and 14 antimicrobial susceptibility data sets were obtained. Ribotyping (11 participants) correctly identified most RTs (median 91% concordance rate) except for RT002, which was misidentified in 4/11 reports. However, this isolate was correctly asserted to RT002 after an update of a publicly available ribotyping database. Multilocus sequence typing, surface layer sequence typing, DNA microarray based genotyping, and whole genome sequencing, which were used by 1–3 participants, identified all six isolates correctly. AST was done by epsilometry by the participants and compared to agar dilution data determined by the coordinating reference centre. Susceptibilities against metronidazole, moxifloxacin, and vancomycin were correctly identified in 235 of 237 cases and in accordance to agar dilution as the gold standard. Genotyping of the C. difficile test strains revealed a remarkable high concordance on the level of ribotypes with a wide variety of methods. Epsilometry appears to be a reliable method for AST of C. difficile isolates in routine clinical microbiology laboratories.

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