Typing and subtyping of Clostridium difficile isolates by using multiple-locus variable-number tandem-repeat analysis.
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.
van den Berg
Renate J
RJ
Department of Medical Microbiology, Center of Infectious Diseases, Leiden University Medical Center, Leiden, The Netherlands.
Schaap
Inge
I
Templeton
Kate E
KE
Klaassen
Corné H W
CH
Kuijper
Ed J
EJ
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
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004269:DNA, Bacterial; D004768:Enterotoxins; C041348:tcdA protein, Clostridium difficile; C031815:toxB protein, Clostridium difficile