Analysis of the genetic distribution among members of Clostridium botulinum group I using a novel multilocus sequence typing (MLST) assay.
Clostridium botulinum is the etiological agent of botulism. Due to food-borne poisoning and the potential use of the extremely toxic botulinum neurotoxin (BoNT) from C. botulinum in bioterror or biocrime related actions, reliable high resolution typing methods for discriminating C. botulinum strains are needed. Partial sequencing of the adk, atpH, gyrB, proC, rpoD and spo0A genes from 51 various C. botulinum/sporogenes isolates was performed, resulting in 37 different sequence types (STs). Analysis of the sequence data revealed a genetic distribution in five larger clusters with a loose correlation to the BoNT serotypes. The developed MLST assay had a slightly lower resolution ability when compared to the MLVA (multilocus variable number of tandem repeat analysis), but the two methods resulted in similar subclusters of the strains possessing the BoNT serotypes A, B and F. The current work presents the development of a novel MLST assay useful for genotyping C. botulinum related to basic phylogenetic research and trace-back analysis in microbial forensic studies.
Norwegian Defence Research Establishment, P.O. Box 25, N-2027 Kjeller, Norway. Electronic address: Jaran-Strand.Olsen@ffi.no.
Jaran S
JS
Olsen
German Armed Forces, Institute of Microbiology, Munich, Germany.
Holger
H
Scholz
Army Medical and Veterinary Research Center, Via Santo Stefano Rotondo 4, I-00184 Rome, Italy.
Silvia
S
Fillo
Division of Analytical Microbiology, DGA CBRN Defence, BP3, 91710 Vert le Petit, France.
Vincent
V
Ramisse
Army Medical and Veterinary Research Center, Via Santo Stefano Rotondo 4, I-00184 Rome, Italy.
Florigio
F
Lista
Norwegian Defence Research Establishment, P.O. Box 25, N-2027 Kjeller, Norway.
Anette K
AK
Trømborg
Norwegian Defence Research Establishment, P.O. Box 25, N-2027 Kjeller, Norway.
Tone
T
Aarskaug
Norwegian Defence Research Establishment, P.O. Box 25, N-2027 Kjeller, Norway.
Ingjerd
I
Thrane
Norwegian Defence Research Establishment, P.O. Box 25, N-2027 Kjeller, Norway.
Janet M
JM
Blatny
D001426:Bacterial Proteins / Q000235:genetics; D003014:Clostridium botulinum / Q000145:classification* / Q000235:genetics*; D016000:Cluster Analysis; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D005838:Genotype; D006801:Humans; D008969:Molecular Sequence Data; D058885:Multilocus Sequence Typing / Q000379:methods*; D010802:Phylogeny; D017422:Sequence Analysis, DNA; D017385:Sequence Homology; D012703:Serotyping
D001426:Bacterial Proteins; D004269:DNA, Bacterial
Clostridium botulinum; MLST; MLVA; Phylogenetic