Clostridium botulinum Group I Strain Genotyping by 15-Locus Multilocus Variable-Number Tandem-Repeat Analysis

Fillo, Silvia
Giordani, Francesco
Fabrizio, Anniballi
Gorgé, Olivier
Ramisse, Vincent
Vergnaud, Gilles
Riehm, Julia
Scholz, Holger C.
Splettstoesser, Wolf
Kieboom, Jasper
Olsen, Jaran Strand
Fenicia, Lucia
Lista, Florigio
Fillo, Silvia; Giordani, Francesco; Fabrizio, Anniballi; Gorgé, Olivier; Ramisse, Vincent; Vergnaud, Gilles; Riehm, Julia; Scholz, Holger C.; Splettstoesser, Wolf; Kieboom, Jasper; Olsen, Jaran Strand; Fenicia, Lucia; Lista, Florigio. Clostridium botulinum Group I Strain Genotyping by 15-Locus Multilocus Variable-Number Tandem-Repeat Analysis. Journal of Clinical Microbiology 2011 ;Volum 49.(12) s. 4252-4263
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Clostridium botulinum is a taxonomic designation that encompasses a broad variety of spore-forming, Gram-positive bacteria producing the botulinum neurotoxin (BoNT). C. botulinum is the etiologic agent of botulism, a rare but severe neuroparalytic disease. Fine-resolution genetic characterization of C. botulinum isolates of any BoNT type is relevant for both epidemiological studies and forensic microbiology. A 10-locus multiple-locus variable-number tandem-repeat analysis (MLVA) was previously applied to isolates of C. botulinum type A. The present study includes five additional loci designed to better address proteolytic B and F serotypes. We investigated 79 C. botulinum group I strains isolated from human and food samples in several European countries, including types A (28), B (36), AB (4), and F (11) strains, and 5 nontoxic Clostridium sporogenes. Additional data were deduced from in silico analysis of 10 available fully sequenced genomes. This 15-locus MLVA (MLVA-15) scheme identified 86 distinct genotypes that clustered consistently with the results of amplified fragment length polymorphism (AFLP) and MLVA genotyping in previous reports. An MLVA-7 scheme, a subset of the MLVA-15, performed on a lab-on-a-chip device using a nonfluorescent subset of primers, is also proposed as a first-line assay. The phylogenetic grouping obtained with the MLVA-7 does not differ significantly from that generated by the MLVA-15. To our knowledge, this report is the first to analyze genetic variability among all of the C. botulinum group I serotypes by MLVA. Our data provide new insights into the genetic variability of group I C. botulinum isolates worldwide and demonstrate that this group is genetically highly diverse.
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