title: Typing and subtyping of Clostridium difficile isolates by using multiple-locus variable-number tandem-repeat analysis.
abstract: Using the genomic sequence of Clostridium difficile strain 630, we developed multiple-locus variable-number tandem-repeat analysis (MLVA) with automated fragment analysis and multicolored capillary electrophoresis as a typing method for C. difficile. All reference strains, representing 31 serogroups, 25 toxinotypes, and 7 known subtypes of PCR ribotype 001, could be discriminated from each other. Application of MLVA to 28 isolates from 7 outbreaks due to the emerging hypervirulent PCR ribotype 027-pulsed-field gel electrophoresis type NAP1 resulted in recognition of 13 clusters. Additionally, 29 toxin A-negative, toxin B-positive isolates belonging to PCR ribotype 017 from eight different countries revealed eight country-specific clusters. MLVA is a highly discriminatory genotyping method and a new tool for subtyping of newly emerging variants of C. difficile.
authors.affiliation: Department of Medical Microbiology, Center of Infectious Diseases, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Renate J
authors.initials: RJ
authors.lastname: van den Berg
authors.forename: Inge
authors.initials: I
authors.lastname: Schaap
authors.forename: Kate E
authors.initials: KE
authors.lastname: Templeton
authors.forename: Corné H W
authors.initials: CH
authors.lastname: Klaassen
authors.forename: Ed J
authors.initials: EJ
authors.lastname: Kuijper
mesh_terms: D001331:Automation; D001426:Bacterial Proteins / Q000378:metabolism; D001427:Bacterial Toxins / Q000378:metabolism; D015373:Bacterial Typing Techniques; D016360:Clostridioides difficile / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D004269:DNA, Bacterial / Q000032:analysis; D019075:Electrophoresis, Capillary / Q000379:methods*; D004761:Enterocolitis, Pseudomembranous / Q000382:microbiology; D004768:Enterotoxins / Q000378:metabolism; D005838:Genotype; D006801:Humans; D018598:Minisatellite Repeats; D021521:Ribotyping
chemical_list: D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004269:DNA, Bacterial; D004768:Enterotoxins; C041348:tcdA protein, Clostridium difficile; C031815:toxB protein, Clostridium difficile