title: DNA-Microarray-based Genotyping of Clostridium difficile.
abstract: BACKGROUND
  Clostridium difficile can cause antibiotic-associated diarrhea and a possibility of outbreaks in hospital settings warrants molecular typing. A microarray was designed that included toxin genes (tcdA/B, cdtA/B), genes related to antimicrobial resistance, the slpA gene and additional variable genes.
  
  
  RESULTS
  DNA of six reference strains and 234 clinical isolates from South-Western and Eastern Germany was subjected to linear amplification and labeling with dUTP-linked biotin. Amplicons were hybridized to microarrays providing information on the presence of target genes and on their alleles. Tested isolates were assigned to 37 distinct profiles that clustered mainly according to MLST-defined clades. Three additional profiles were predicted from published genome sequences, although they were not found experimentally.
  
  
  CONCLUSIONS
  The microarray based assay allows rapid and high-throughput genotyping of clinical C. difficile isolates including toxin gene detection and strain assignment. Overall hybridization profiles correlated with MLST-derived clades.
authors.affiliation: Institute for Medical Microbiology and Hygiene, Technische Universität Dresden, Dresden, Germany. darius.gawlik@web.de.|Hamm-Lippstedt University, Hamm, Germany. darius.gawlik@web.de.
authors.forename: Darius
authors.initials: D
authors.lastname: Gawlik
authors.affiliation: Alere Technologies GmbH, Jena, Germany. piet@clondiag.com.|Infectognostics Research Campus, Jena, Germany. piet@clondiag.com.
authors.forename: Peter
authors.initials: P
authors.lastname: Slickers
authors.affiliation: Alere Technologies GmbH, Jena, Germany. ines@clondiag.com.|Infectognostics Research Campus, Jena, Germany. ines@clondiag.com.
authors.forename: Ines
authors.initials: I
authors.lastname: Engelmann
authors.affiliation: Alere Technologies GmbH, Jena, Germany. elke@clondiag.com.|Infectognostics Research Campus, Jena, Germany. elke@clondiag.com.
authors.forename: Elke
authors.initials: E
authors.lastname: Müller
authors.affiliation: Institute for Medical Microbiology and Hygiene, Technische Universität Dresden, Dresden, Germany. christian.lueck@tu-dresden.de.
authors.forename: Christian
authors.initials: C
authors.lastname: Lück
authors.affiliation: Department of Internal Medicine I, University Hospital Schleswig-Holstein, Campus Kiel, Kiel, Germany. anette.friedrichs@uksh.de.
authors.forename: Anette
authors.initials: A
authors.lastname: Friedrichs
authors.affiliation: Alere Technologies GmbH, Jena, Germany. ralf@clondiag.com.|Infectognostics Research Campus, Jena, Germany. ralf@clondiag.com.
authors.forename: Ralf
authors.initials: R
authors.lastname: Ehricht
authors.affiliation: Institute for Medical Microbiology and Hygiene, Technische Universität Dresden, Dresden, Germany. monecke@rocketmail.com.|Alere Technologies GmbH, Jena, Germany. monecke@rocketmail.com.|Infectognostics Research Campus, Jena, Germany. monecke@rocketmail.com.
authors.forename: Stefan
authors.initials: S
authors.lastname: Monecke
mesh_terms: D016360:Clostridioides difficile / Q000145:classification* / Q000235:genetics*; D016000:Cluster Analysis; D005798:Genes, Bacterial; D005838:Genotype; D060005:Genotyping Techniques / Q000379:methods*; D005858:Germany; D057166:High-Throughput Screening Assays; D046228:Microarray Analysis / Q000379:methods*; D058889:Molecular Typing / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques; D009693:Nucleic Acid Hybridization; D013194:Staining and Labeling / Q000379:methods; D013997:Time Factors