Multilocus variable-number tandem-repeat analysis and multilocus sequence typing reveal genetic relationships among Clostridium difficile isolates genotyped by restriction endonuclease analysis.
Numbers of Clostridium difficile infections have increased worldwide in the past decade. While infection with C. difficile remains predominantly a health care-associated infection, there may also be an increased incidence of community-associated infections. C. difficile strains of public health significance continue to emerge, and reliable genotyping methods for epidemiological investigations and global surveillance of C. difficile are required. In this study, multilocus sequence typing (MLST) and multilocus variable-number tandem-repeat analysis (MLVA) were performed on a set of 157 spatially and temporally diverse C. difficile isolates that had been previously genotyped by restriction endonuclease analysis (REA) to determine the concordance among these genotyping methods. In addition, sequence analysis of the tcdC genotype was performed to investigate the association of allelic variants with epidemic C. difficile isolates. Overall, the MLST and MLVA data were concordant with REA genotyping data. MLST was less discriminatory than either MLVA or REA, yet this method established C. difficile genetic lineage. MLVA was highly discriminatory and demonstrated relationships among the MLST genetic lineages and REA genotypes that were previously unrecognized. Several tcdC genotypes were specific to epidemic clones, highlighting the possible importance of toxin misregulation in C. difficile disease pathogenesis. This study demonstrates that a combination of MLST and MLVA may prove useful for the investigation and surveillance of emergent C. difficile clones of global public health concern.
Marsh
Jane W
JW
Infectious Diseases Epidemiology Research Unit, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA. jwmarsh@pitt.edu
O'Leary
Mary M
MM
Shutt
Kathleen A
KA
Sambol
Susan P
SP
Johnson
Stuart
S
Gerding
Dale N
DN
Harrison
Lee H
LH
D001426:Bacterial Proteins / Q000235:genetics; D015373:Bacterial Typing Techniques / Q000379:methods*; D016360:Clostridioides difficile / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D003015:Clostridium Infections / Q000382:microbiology*; D016000:Cluster Analysis; D016172:DNA Fingerprinting / Q000379:methods*; D004269:DNA, Bacterial / Q000235:genetics; D005838:Genotype; D006801:Humans; D018598:Minisatellite Repeats; D017720:Molecular Epidemiology / Q000379:methods; D011110:Polymorphism, Genetic; D000091402:Prohibitins; D012097:Repressor Proteins / Q000235:genetics; D015183:Restriction Mapping; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
D001426:Bacterial Proteins; D004269:DNA, Bacterial; C000717331:PHB2 protein, human; D000091402:Prohibitins; D012097:Repressor Proteins; C105615:TcdC protein, Clostridium difficile